SUPPLEMENTARY ONLINE MATERIAL
Spaceflight-related suboptimal conditions can accentuate Drosophila
genome altered gravity response
R. Herranz, A. Benguría, D. A. Laván, I. López- Vidriero, G. Gasset, F. J. Medina, J. J.W.A. van Loon & R. Marco.
This supplement contains:
Materials and Methods
1. Constraints of a space experiment.
2. Detailed description of the space experiment.
3. The ground simulation equipment: the Random Position Machine and the
hypergravity centrifuge.
4. Microarray analysis.
5. GeneOntology analyses.
6. Quantitative PCR validation of the Microarray data.
7. GEDI and Pathway Studio analyses.
Figures S1 to S6
Tables S1 & S2 (as a separate excel file).
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1. Constraints of a space experiment
Spaceflight experiment poses inherent limitation unlike regular ground based research. To recall
only the more critical ones: 1. The launch capability in the Soyuz capsule is quite limited both in
volume and in weight. 2. Although several research modules have been uploaded to the ISS not all
of them were available at the time of our study (end 2003). For example, no glovebox was available
for biological experiments, complicating the fixation and manipulation steps of our samples. 3. The
main objective of the missions this experiment was part of, was the replacement of two or three ISS
crew members. (J.J. van Loon, F.J. Medina and R. Marco. Microgravity Sci Technol 19, 9-32 (2007)).
In this particular mission (Cervantes) astronaut Pedro Duque performed our and other experiments
linked this mission. The rest of the crew provides assistance, if they had time available, a scarce
resource on the ISS. 4. The complete flight is quite short, around 10 days. During ascent and
descent parts of the mission, the samples were exposed to the same environmental as the astronauts.
There was no active temperature control during these phases. A temperature logger allowed to
retrospectively controlling the temperature profile from the mission. 5. The laboratories at the
launch site Baikonur were still primitively equipped. Thus, the experiments had to be transported
from the original laboratories to the launching site already prepared, ready for launch. Being
Baikonur in a different country this transport step has to be organized and took four days to prevent
missing the launch if any delay occurred. The experiment had to be stabilized or delayed for this
logistical constraint. 6. The landing place is only approximately known. It takes some time to reach
it using helicopters and there is a bog limitation for investigators to be present at the landing site to
process their samples as soon as possible. Transport to Star City, near Moscow, took 12 hours for
this mission. Laboratory facilities in Star City were primitive. This constraint causes a low yield
RNA extraction but we established in our preliminary report of the GENE experiment that RNA
quality was good enough to be used in the microarray analysis (R. Herranz et al., J Gravit Physiol
12, 1, 253-254 (2005)).
2. The Space experiment
It took four days to move our samples from Western Europe to the launching site at Baikonur in
Kazakhstan. As can be deduced from the temperature recording reproduced in Figure S1, the
samples were transported at 14ºC until the handover took place in Baikonour. We had previously
found that Drosophila larvae and pupae are able to survive when exposed to relatively low
temperatures simply slowing down the larval and pupal development (P. Anthony, J. Ausseil, B.
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Bechler, A. Benguría, N. Blackhall et al. J. Biotechnol. 47, 377-393 (1996)). The air inside the
hermetical containers is sufficient to allow complete development of the pupae under these
conditions (R. Herranz, D.A. Laván, F.J. Medina, J.J. van Loon, R. Marco. Microgravity Sci.
Technol 21, 4, 299-304 (2009)). It is known from experiments in the 1940s (A. Wolsky. Science.
94, 2428, 48-49 (1941); B. Sacktor. J Gen Physiol. 35, 3, 397-407 (1952); D. Bodenstein, B.
Sacktor. Science. 116,3012, 299-300 (1952)) that the oxygen consumption during pupal
development decreases significantly at the beginning of pupal development and increases again at
eclosion. Limiting the oxygen simply slows down the hatching of the imagoes from the pupae at the
end of pupal development without affecting their survival. The sample containers containing the
pupae were handed over to the ESA representatives the evening before the Soyuz launch on 10 Oct.
2003 (Fig. S2). After docking to the ISS some 50 hrs. after launch, the samples were transferred to
the Aquarius B incubator set at 22ºC in the Russian Segment of the ISS. Thus, after three days and a
half in microgravity crew took them out of the incubator and activate the acetone fixation step (Fig.
S3). Quickly, the containers were stored in the Kryogen freezer at -22ºC until the end of the
mission, when they were taken out and installed back into the same Biology Transport Container
(BTC) that had been used to bring them to the ISS. Once landed the containers in the BTC were
transferred to a cold transport box set at 3ºC and delivered to us at Star City in Moscow where the
containers were opened, and the RNA extracted from the fixed pupae (see below). The RNAs were
further purified and used for microarray analysis in Madrid. The adaptation of this methodology to
the space constraints has been described (Herranz, R. D. Husson, A. Villa, M. Pastor, F. J. Medina
& R. Marco. J Gravit Physiol. 12, 2, 51-60 (2005)).
The pupae grown in Madrid, prepared, inside double containment gas permeable plastic bags and
loaded in Toulouse some 4 days before launch. The bags contained a filter paper with 20 white
Drosophila pupae, collected from the walls of plastic tubes as they were initiating pupation. Inside
the plastic bags (Fig. S3A), two sets of symmetrically located ampoules filled with 80 l each of
pure dehydrated acetone each are placed below the filter paper. Finally, two bags are mounted in an
internal frame with two pestles that can be activated independently to break each set of ampoules at
a preset time. Acetone was used as an RNA preservative together with a below -20ºC temperature
during 6 days, a procedure previously tested by us (Herranz, R. D. Husson, A. Villa, M. Pastor, F. J.
Medina & R. Marco. J Gravit Physiol. 12, 2, 51-60 (2005)).
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A power unit (Fig. S3C) able to activate the motor was sent previously within a Progress
resupplying flight to the ISS. The whole mounting is inserted in an ESA type-I/E container 2x4x8
cm. In the original Berlingot concept (M. Ayed, O. Pironneau O, H. Planel H, G. Gasset G, G.
Richoilley G. Microgravity Sci. Technol. 5, 2, 98-102 (1992)), the operations was manual and
required a glovebox. However, since no glovebox was available to us at the time of our experiment,
we used the so-called MAMBA concept developed by Dutch Space (Leiden, the Netherlands, Fig.
S1B). In the MAMBA a tiny electrical motor activates the pestles when powered at a preset time.
The adaptation of this methodology has been described elsewhere (Herranz, R. D. Husson, A. Villa,
M. Pastor, F. J. Medina, R. Marco. J Gravit Physiol. 12, 2, 51-60, (2005)). Soft foam transport
boxes were used to carry the containers to the ISS in the Soyuz space craft and back (Fig. S2).
3. The ground microgravity simulation and the hypergravity centrifuge. The RPM and the
MidiCAR centrifuge
Information on these instruments can be found in the following Internet site: http://www.descsite.nl
The Random Positioning Machine (RPM) is a microweight ('microgravity') simulator that is based
on the principle of 'gravity-vector-averaging'. The system may be compared with a classic 2D
clinostat although such a clinostat has only a two dimensional averaging of the g vector while the
RPM
provides
a
functional
volume
which
is
'exposed'
to
simulated
microweight.
Gravity is a vector, i.e. it has a magnitude and a direction. During an experiment run in this two axis
RPM the sample's position with regard to the Earth's gravity vector direction is constantly changing.
The sample may experience this as a zero-gravity environment (Fig. S4A; See Van Loon, J.J.W.A.
Adv. Space Res. 39, 1161-5(2007) for additional details).
The Medium Sized Centrifuge for Acceleration Research (MidiCAR) is a dedicated centrifuge in
which samples may be exposed to accelerations up to 100 times Earth's gravity. The facility is
accommodated in a temperature-controlled incubator and driven by dedicated software. Gravity
levels may be chosen according specific user requirements, either static or dynamic. The system
may be applied for short term (seconds) or long term (weeks) studies. Dedicated experiment vessels
are compatible with standard laboratory tissue / cell culture hardware but also flight specific
modules may be accommodated. Both rotating and static control samples are housed in the same
environment (Fig. S4B; J.J.W.A. van Loon, L.C. van den Bergh, R. Schelling, J.P. Veldhuijzen,
R.H. Huijser. 44th International Astronautical Congress, IAF/IAA-93-G.4.166, Gratz, Austria,
October 1993).
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A series of constrained (as similar as possible to the Soyuz mission timelines, temperatures and
containers) and non constrained experiments were performed in the RPM. Only non constrained
experiments were done in the MidiCAR centrifuge at a 10g effective force.
4. Microarray analysis
Total RNA was extracted from 15 to 20 late pupa on each filter paper recovered from the flight
using TRIZOL reagent (reference number 15596-026, Invitrogen lifescience technologies). RNA
pellets were conserved with supernatant ethanol and transported to our lab in Spain on dry ice.
Then, RNA was raised on DEPC water and further purified with RNeasy mini kit (QIAGEN).
Purified RNAs were biotin-labeled using the One cycle target-labeling kit (Affymetrix, Santa Clara,
CA). cDNA was synthesized from 5 g total RNA using an oligo-dT primer with a T7 RNA
polymerase promoter site added to the 3´ end. After second-strand synthesis, in vitro transcription
was performed using T7 RNA polymerase to produce biotin labeled cRNA. The cRNA preparations
(20 g) were fragmented at 95oC for 35 min into 35-200 bases length and added to a hybridization
solution containing 100 mmol/l 2-(N-morpholino)ethanesulfonic acid, 1 mol/l Na+ and 20 mmol/l
of EDTA in the presence of 0.01% Tween 20 to a final cRNA concentration of 0.05 g/ml.
5 g of cRNA were hybridized to Test3 arrays (Affymetrix) for sample quality control. 15 g of
cRNA were then hybridized to the Drosophila Genome Array (Affymetrix).
Each microarray was washed and stained in a Fluidics station 400 (Affymetrix) following the
standard protocol and scanned at 3 µm resolution in an Agilent HP G2500A GeneArray scanner
(Agilent Technologies). A total of 20 CEL files have been obtained from the hybridized arrays. The
replicate samples of the experiments when clustered are coherent (Fig. S5) but it is difficult to
perform inter-experiment comparisons since the 1g controls in the non constrained experiments are
not comparable but the constrained 1g controls are (indicating a good correlation of the ISS and the
RPM simulation).
Data analysis was performed using the affylmGUI R package (J.M. Wettenhall, K.M. Simpson, K.
Satterley, G.K. Smyth. Bioinformatics. 22, 7, 897-899 (2006)). Robust Multi-array Analysis (RMA)
algorithm was used for background correction, normalization and expression levels summarization
(R.A. Irizarry, B. Hobbs, F. Collin, Y.D. Beazer-Barclay,K.J. Antonellis et al. Biostatistics. 4, 2,
249-64, 2003). Next, differential expression analysis was performed using the Bayes t-statistics
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from the linear models for microarray data (limma), included in the affylmGUI package. We have
chosen the commonly used Benjamini-Hochberg’s method for p-value correction False Discovery
Rates available in the affylma R package because it is one of the most widely used and accepted in
microarray experiments and it is implemented in the affylma R package for p-value correction after
running Bayes t-statistic analysis (Y. Benjamini, Y. Hochberg. J.Roy.Stat.So. 57, 289-300 (1995);
A. Reiner, D. Yekutieli, Y. Benjamini. Bioinformatics. 19, 3, 368-375 (2003); Storey J.D. J. R. Stat.
Soc. B 64, 479–498 (2002)). Venn diagrams were obtained using the Venny program (Oliveros, J.C.
VENNY.
An
interactive
tool
for
comparing
lists
with
Venn
Diagrams.
http://bioinfogp.cnb.csic.es/tools/venny/index.html (2007)). Alternatively, MAS 5.0 algorithm
(Affymetrix) was used for normalization and summarization of the arrays. The original data has
been submitted to the EMBL European Bioinformatic Institute for Microarray Informatics using the
MIAMExpress annotation tool following the Miame recommendations and will be publicly released
when accepted for publication.
5. GeneOntology Analysis
Gene Ontology enrichment analysis of overlapping DEGs found in the ISS and RPM constrained
experiments was performed using DAVID Functional Annotation tool (D. Huang, B. Sherman, R.
Lempicki. Nature Protoc. 4, 1, 44-57 (2009)). Raw p-values were corrected using the Benjamini
Hochsberg´s method and terms with p-value < 0.05 were selected.
6. Quantitative RT-PCR analysis
Figure 2B (in the main article) shows the confirmation analysis that compares the signal log ratio
difference between a defined set of genes obtained by microarrays and by quantitative realtime PCR
under the conditions described hereafter. The fluorescent probes UPL and the primers were
designed and selected using the Roche Universal Probe Library software for Real-Time qPCR
assays, as recommended by the manufacturer.
For the retrotranscription: 1 µg RNA per 100 µl reaction (or 500 ng/ 50 µl) using the High Capacity
Archive kit of Applied Biosystems: 10 µl 10x buffer, 10 µl Random Primers 10x, 4 µl dNTPs 25x
and 5 µl Multiscribe 50 U/µl. Incubations were 10 min at 25ºC and 120 min at 37ºC.
For the quantitative PCR using UPL probes in (reacción 10 µl reaction volume, using the Applied
Biosystems Abi Prism 7.900HT equipment): 5 µl 2x TaqMan MasterMix (Roche), 0,1 µl probe 10
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µM (final concentration 100 nM), 0,2 µl oligo F 12,5 µM (final concentration 250 nM), 0,2 µl oligo
R 12,5 µM (final concentration 250 nM), 0,5 µl cDNA (10 ng/µl; final concentration 0,5 ng/ µl) and
4 µl de Nuclease-free H2O (Ambion). The eukaryotic 18S rRNA probe was purchased from AB.
Conditions of PCR: 2 min at 50 ºC,10 min at 95 ºC, 40 cycles of 15 sec at 95 ºC followed by 1 min
@60ºC RQ, relative quantities of transcripts were calculated as follows. Expressions levels relative
to a reference sample, both normalized to a housekeeping gene (18S rRNA or RNA Pol-II).
RQ = 2 –ΔΔCt where for each sample, ΔΔCt = ΔCt sample – ΔCt ref and ΔCt = Ctgen – Ctendo.
7. GEDI and Pathway Studio analyses
GEDI analysis was done using version 2.1 of the software. First, we applied the RMA algorithm for
background correction, normalization and expression level summarization of the arrays (see above)
using GeneSpring v10 software (Agilent Technologies). Probe sets with expression level below 20 th
percentile in 50% of conditions were filtered out, leaving 10995 probe sets. Next, we calculated the
average log2ratios for each experimental condition versus 1g control, and used this value for GEDI
analysis. Mosaics of 15 x 13 grid size (average of 56 genes/tile) were obtained using the following
settings of the software:
Training iterations:
first phase: 60
second phase: 120
Neighborhood radius:
first phase: 4.0
second phase: 1.0
Learning factor:
first phase: 0.5
second phase: 0.05
Neighborhood block size:
first phase: 4
second phase: 2
Conscience:
first phase: 3.0
second phase: 3.0
Random seed: 1
Distance metrics: Euclidean Linear Initialization Method
Pathway Studio demo version 6.1 (Ariadne Genomics) was used to find the relationships among a
number of genes that share a significant change in their expression level in at least three of the four
conditions analyzed + a virtual fifth condition (pooled analysis of the constrained experiment
justified by the clustering of the ISSc and RPMc conditions in Figure S5 tree). These results are
included in Figure S6 and enlisted in table S2.
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FIGURE LEGENDS:
Figure S1: Diagram of the temperature profile and time line of the ISS experiment. Transport from
Madrid, Spain (M), Preparation of the pupae and insertion of the Gene containers in the transport
container in Toulouse, France (T), handover of the sample in Baikonur / Kazakhstan (B), launch of the
Soyuz rocket (L), arrival in the ISS (I) and fixation & transfer to the Kryogen Freezer (F).
Figure S2. The BTU (Biological Transport Unit). Soft foam transport boxes used to take the samples
up to and down from the ISS in the Soyuz space craft. The BTU did not provide an active temperature
control
Figure S3. Pictures of the spaceflight hardware. The sample setting (A). Each double layered bag
contained 20 pupae and 960 µl of acetone in glass ampoulae. The MAMBA hardware (B) and the Power
Unit (C). MAMBA outer view, inner view, with sample bag (berlingot) in position and partially inserted
in ESA standard Type-I/E experiment container (internal dimensions: 8x4x2 cm).
Figure S4. The Random Position Machine and MidiCAR centrifuge from DESC. A Random
Position Machine is an instrument made up by two frames that are able to rotate independently,
allowing the sample to adopt all the different positions with respect to the gravity vector. See for
additional details: Van Loon, J.J.W.A. Adv. Space Res, 39, 1161-1165 (2007) and A. G. Borst, J.J.W. A.
van Loon. Microgravity Sci. Technol, 21, 287-292 (2009). The MidiCAR is a dedicated centrifuge for
cell and tissue culture experiments at moderate hypergravity levels: 1-100xg. (van Loon J.J.W.A., L.C.
van den Bergh, R. Schelling, J.P. Veldhuijzen, R.H. Huijser. 44th International Astronautical Congress,
IAF/IAA-93-G.4.166, Gratz, Austria, October 1993).
Figure S5. Hierarchical clustering of the different experiments analyzed in this study and
accession numbers.
HG, hypergravity, RPM, Random Position Machine, ISS, International Space Station. A) All branches
have a 100% confidence using Pearson correlation algorithm with 100 iterations (Genespring software).
The distance indicates the number of iterations in which the branches collapse. It shows the clustering
tree among the expression data arrays. RPM and ISS data cluster together suggesting a very good
correlation between simulated and real microgravity conditions with constraints. B) Table including the
description of the 21 CEL files used in this paper is shown below. From then, 20 has been used in
the main analysis, while one does not fit with the quality control standards and it is not consistent
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with other raw data files. The whole experiment has been submitted to the MIAMExpress database
with the accession number E-MEXP-2064.
Figure S6: Relations among the 43 genes with a remarkable change in expression (2 fold) in at
least three of the analyzed conditions. Main image illustrate the expression level of the 43 genes in the
pooled constrained analysis (ISS plus RPM, included as the fifth condition for this analysis) as log
change vs. 1g constrained controls. The genes used for the connection have been shaded in order to
highlight the affected genes (upregulated in red, downregulated in blue). The kind of proteins/processes
involved (when available in the database) and the type of relation between the entities are indicated in
the legend. Four small insets corresponding to same genes expression levels in the non pooled
conditions (ISS and RPM with the container constraints and RPM and 10g experiment without the
container constraints) are included. Excel Table S2 with their expression values is available.
Table S1. Gene Ontologies (GO) significantly up regulated or down regulated of the overlapping DEGs
on real or simulated microgravity (ISS/RPM) under constrained conditions.
Table S2. 50 Probesets signal and p values from the 43 candidate genes that arose from the Figure S6
pathway analysis (Included as a separate excel file).
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Temperature (ºC)
FIGURE S1
Date
FIGURE S2
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FIGURE S3
A)
B)
C)
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FIGURE S4
A)
B)
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Page S.12
FIGURE S5
A)
HG experiment (10g)
Distance 0.01
HG control (1g)
Without
constraints
RPM experiment (sim. µg)
RPM control (1g)
RPM control (1g)
ISS control (1g)
With
constraints
RPM experiment (sim. µg)
ISS experiment (µg)
B)
Experiment details
Gravity facility
ISS
8 days with transport (cold) &
containment (hipoxia)constraints
Condition
Sample name in E-MEXP-2064
0g (not used)
Rmarco_Droso_FG1
0g
1g
0g
RPM
1g
RMarco_Droso_FG3
RMarco_Droso_FG4
RMarco_Droso_GG4
RMarco_Droso_GG5
RMarco_ Droso_5
RMarco_Droso_7
RMarco_Droso_81
RMarco_Droso_82
RMarco_Droso_91
0g
RMarco_Droso_92
RMarco_Droso_101
RPM
RMarco_Droso_112
1g
5 days without transport (cold) &
containment (hipoxia) constraints
RMarco_Droso_121
RMarco_Droso_122
RMarco_Droso_C1
1g
RMarco_Droso_F1
RMarco_Droso_F2
Centrifuge
(hypergravity)
RMarco_Droso_D1
10g
RMarco_Droso_D2
RMarco_Droso_E2
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FIGURE S6
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TABLE S1
GO_CellularComponent_UP
Term
Corr p-value
GO:0044455~mitochondrial membrane part
3.32E-38
GO:0005746~mitochondrial respiratory chain
3.35E-36
GO:0044429~mitochondrial part
2.15E-34
GO:0005743~mitochondrial inner membrane
1.27E-33
GO:0019866~organelle inner membrane
4.16E-33
GO:0031966~mitochondrial membrane
7.18E-32
GO:0005740~mitochondrial envelope
7.86E-32
GO:0005739~mitochondrion
2.18E-30
GO:0031967~organelle envelope
3.54E-27
GO:0031975~envelope
4.08E-27
GO:0031090~organelle membrane
1.88E-21
GO:0030964~NADH dehydrogenase complex (quinone)
1.95E-21
GO:0005747~mitochondrial respiratory chain complex I
1.95E-21
GO:0045271~respiratory chain complex I
1.95E-21
GO:0044444~cytoplasmic part
6.20E-17
GO:0005737~cytoplasm
4.96E-16
GO:0032991~macromolecular complex
3.77E-12
GO:0044446~intracellular organelle part
1.16E-11
GO:0044422~organelle part
1.19E-11
GO:0043234~protein complex
4.92E-11
GO:0044425~membrane part
1.53E-08
GO:0005750~mitochondrial respiratory chain complex III
8.37E-06
GO:0045275~respiratory chain complex III
8.37E-06
GO:0016020~membrane
3.00E-05
GO:0005753~mitochondrial proton-transporting ATP synthase complex
2.30E-04
GO:0044449~contractile fiber part
3.48E-04
GO:0045259~proton-transporting ATP synthase complex
3.50E-04
GO:0043292~contractile fiber
6.66E-04
GO:0005762~mitochondrial large ribosomal subunit
7.84E-04
GO:0000315~organellar large ribosomal subunit
7.84E-04
GO:0043229~intracellular organelle
9.42E-04
GO:0043226~organelle
9.45E-04
GO:0043231~intracellular membrane-bound organelle
0.00101436
GO:0043227~membrane-bound organelle
0.00103282
GO:0005751~mitochondrial respiratory chain complex IV
0.00119393
GO:0045277~respiratory chain complex IV
0.00119393
GO:0005761~mitochondrial ribosome
0.00120956
GO:0000313~organellar ribosome
0.00120956
GO:0000275~mitochondrial proton-transporting ATP synthase complex, catalytic core F(1)
0.00326251
GO:0005759~mitochondrial matrix
0.00330117
GO:0031980~mitochondrial lumen
0.00330117
GO:0030017~sarcomere
0.00428427
GO:0030016~myofibril
0.00428427
GO:0045261~proton-transporting ATP synthase complex, catalytic core F(1)
0.00556587
GO:0042995~cell projection
0.00701846
GO:0044424~intracellular part
0.01148412
GO:0005865~striated muscle thin filament
0.02536593
GO:0015934~large ribosomal subunit
0.02959205
GO:0016469~proton-transporting two-sector ATPase complex
0.0460202
GO:0009288~flagellin-based flagellum
0.0486683
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GO_Biological process_UP
Term
Corr p-value
GO:0006119~oxidative phosphorylation
3.75E-37
GO:0042775~organelle ATP synthesis coupled electron transport
5.27E-35
GO:0042773~ATP synthesis coupled electron transport
9.11E-35
GO:0006091~generation of precursor metabolites and energy
4.21E-26
GO:0006118~electron transport
1.35E-22
GO:0006120~mitochondrial electron transport, NADH to ubiquinone
3.04E-21
GO:0016310~phosphorylation
1.63E-18
GO:0006796~phosphate metabolic process
5.66E-16
GO:0006793~phosphorus metabolic process
5.66E-16
GO:0006122~mitochondrial electron transport, ubiquinol to cytochrome c
5.37E-05
GO:0006732~coenzyme metabolic process
0.0011638
GO:0051186~cofactor metabolic process
0.00235686
GO_Molecular Function_UP
Term
Corr p-value
GO:0003954~NADH dehydrogenase activity
1.49E-20
GO:0016651~oxidoreductase activity, acting on NADH or NADPH
3.62E-20
GO:0016655~oxidoreductase activity, acting on NADH or NADPH, quinone or similar compound as acceptor
5.22E-17
GO:0050136~NADH dehydrogenase (quinone) activity
5.22E-17
GO:0008137~NADH dehydrogenase (ubiquinone) activity
5.22E-17
GO:0016491~oxidoreductase activity
9.21E-16
GO:0009055~electron carrier activity
1.05E-14
GO:0015078~hydrogen ion transmembrane transporter activity
2.59E-10
GO:0015077~monovalent inorganic cation transmembrane transporter activity
3.00E-10
GO:0022890~inorganic cation transmembrane transporter activity
5.05E-09
GO:0008121~ubiquinol-cytochrome-c reductase activity
7.97E-04
GO:0016681~oxidoreductase activity, acting on diphenols and related substances as donors, cytochrome as acceptor
7.97E-04
GO:0016679~oxidoreductase activity, acting on diphenols and related substances as donors
0.00269371
GO:0003824~catalytic activity
0.003405433
GO:0008324~cation transmembrane transporter activity
0.006468193
GO:0015002~heme-copper terminal oxidase activity
0.007662691
GO:0016676~oxidoreductase activity, acting on heme group of donors, oxygen as acceptor
0.007662691
GO:0016675~oxidoreductase activity, acting on heme group of donors
0.007662691
GO:0004129~cytochrome-c oxidase activity
0.007662691
GO:0015075~ion transmembrane transporter activity
0.00832368
GO:0008553~hydrogen-exporting ATPase activity, phosphorylative mechanism
0.024156962
GO:0015662~ATPase activity, coupled to transmembrane movement of ions, phosphorylative mechanism
0.030225092
GO:0022892~substrate-specific transporter activity
0.037776843
GO:0046961~hydrogen ion transporting ATPase activity, rotational mechanism
0.03804815
GO:0046933~hydrogen ion transporting ATP synthase activity, rotational mechanism
0.03804815
RUNNING TITLE: Microgravity transcriptional profile in Drosophila (Supplementary Material)
Page S.16
GO_CellularComponent_DOWN
Non significant groups
GO_Biological Process_DOWN
Term
Corr p-value
GO:0032502~developmental process
3.15E-06
GO:0007275~multicellular organismal development
6.90E-06
GO:0048731~system development
6.25E-05
GO:0048869~cellular developmental process
7.40E-05
GO:0048856~anatomical structure development
7.78E-05
GO:0030154~cell differentiation
1.25E-04
GO:0048468~cell development
0.001125396
GO:0032501~multicellular organismal process
0.001386307
GO:0006457~protein folding
0.004019878
GO:0048513~organ development
0.006599523
GO:0002165~instar larval or pupal development
0.016325996
GO:0009791~post-embryonic development
0.022581008
GO:0016265~death
0.023076101
GO:0008219~cell death
0.023847463
GO:0016043~cellular component organization and biogenesis
0.02700509
GO_Molecular Function_DOWN
Term
Corr p-value
GO:0005515~protein binding
1.05E-05
GO:0005488~binding
0.04365502
GO:0051082~unfolded protein binding
0.048480782
RUNNING TITLE: Microgravity transcriptional profile in Drosophila (Supplementary Material)
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