Development 143: doi:10.1242/dev.134270: Supplementary information
p72
-Gal4>Cdk1WT-VFP or neur
yw
(n=80)
neur p72-Gal4/+;
UAS-Cdk1WT/+
(n=80)
100%
100%
>1 socket/ 1
shaft
0%
0%
63%
1 socket/
>1 shaft
0%
0%
6%
>1 socket/
>1 shaft
0%
0%
11%
>1 socket/ 0
shaft
0%
0%
6%
1 socket/
1 shaft
p72
-
neur p72-Gal4/+;
UAS-Cdk1F/+
(n=96)
14%
Development • Supplementary information
Table S1. Percentage of macrochaetae defects observed in neur
Gal4>Cdk1F-VPF-expressing adult flies.
Development 143: doi:10.1242/dev.134270: Supplementary information
yw
(n=7)
neur p72-Gal4/+;
UAS-Cdk1WT/+
(n=9)
neur p72-Gal4/+;
UAS-Cdk1F/+
(n=12)
# of short
bristle/adult
notum
229±19
224±11
198±9
1 socket/
1 shaft
100%
100%
55±6.8%
>1 socket/ 1
shaft
0%
0%
26.2±4.2%
1 socket/
>1 shaft
0%
0%
10.8±4.7%
>1 socket/
>1 shaft
0%
0%
4.8±1.4%
>1 socket/ 0
shaft
0%
0%
2.6±1.6%
Development • Supplementary information
Table S2. Percentage of microchaetae defects observed in neur p72-Gal4>Cdk1WT-VFP or neur-Gal4>Cdk1FVFP-expressing adult flies
Fig. S1. Each neuron is associated with a single sheath cell in lineages expressing Cdk1F. Sensory cells
expressing neurP72>Cdk1WT or Cdk1F at 24 hour APF (green). Sheath cells were detected by accumulation of
Prospero (red) and neurons by ELAV (blue). (A – A’’) Cdk1WT-expressing SO has one sheath cell and one
neuron at 24 hours APF. (B – B’’) Lineages expressing Cdk1F also contained one sheath and one neuron cell at
24 hours APF. Note sensory organs with more that the normal set of four sensory cells resulting from extramitosis (arrow).
Development • Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Fig. S2. Ectopic expression of Cdk1WT did not modify the pattern of cell divisions in the microchaetae
lineage. Combined 4D-imaging in living pupae (A-J) of neur p72-Gal4 driven expression of H2B::RFP,
Pon::GFP. (A-J) Representative frames from a time-lapse recording are shown for one cluster and each frame
was temporally localized in the cell lineage schematic shown in panel K for clarity. Color code: pI-cells in blue,
pIIb and its progeny in green, pIIa and its progeny in red. Note that the Pon::GFP reporter is asymmetrically
segregated at each cell division. (K) Schematic representation of cell divisions of this lineage. Note that the final
cluster is composed of 4 cells. The glial cell fragments in panels H and I.
Development • Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Fig. S3. Cdk1F expression in the microchaete lineage forced pI-cells to undergo asymmetric self-renewal
division. Combined 4D-imaging in living pupae expressing neur p72-Gal4 driven expression of Cdk1F and
H2B::RFP, Pon:GFP. (A-L) Representative frames from a time-lapse recording are shown for one cluster and
each frame was temporally localized in the cell lineage schematic shown in M, for clarity. Note that brightly
labeled Pon::GFP is asymmetrically segregated at each division. (M) Schematic representation of cell divisions
of this lineage. Note that the cluster is composed of 8 cells and the anterior pI daughter cell recapitulates the
same pattern of cell divisions as the pI control, resulting in five cells: the two inner cells, a glial cell that enters
in apoptosis, a shaft and a socket cells. (n=59; 62% of lineages were WT and 38% showed self renewal as
depicted (M)). In approximately half of the clusters and additional mitosis in the pIIa sub-lineage was observed,
as shown. (A-M) Color code: pI and anterior pI daughter cells in blue, posterior pI daughter cell and its progeny
in ochre, pIIb and its progeny in green and pIIa and its progeny in rouge.
Development • Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Fig. S4. Ectopic expression of Cdc25stg forced pI-cells to undergo asymmetric self-renewal division in the
microchaete lineage. Combined 4D-imaging in living pupae of neur p72-Gal4 driven expression of Cdc25string,
H2B::RFP, Pon::GFP. (A-H) Representative frames from a time-lapse recording are shown for one cluster and
temporally localized during the cell lineage in I. For clarity, schematic representations are depicted below each
frame. Note that Pon::GFP is asymmetrically segregated at each division. (A-H) color code as in Figure S3. (I)
Shows the schematic representation of cell divisions of this lineage. Note in panels F-H that cells originated
from the posterior pI daughter cell, since does not inherit Pon/Numb, fragments.
Development • Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Development • Supplementary information
Fig. S5. Cells identity after Cdc25stg over-expression. (A-G) Combined 4D live imaging and lineage analysis.
(A-F) Representative frames from a time-lapse recording of one clusters expressing neurp72>Cdc25stg,
H2B::YFP. Note in frame F the fragmentation of posterior pI daughter cells (star). Colour code: pI-cell in blue,
pIIb and its progeny are outlined in green, pIIa cells and its progeny in ochre and secondary pIIa cells and its
progeny in red. (G) Immunostaining of the same cluster after the completion of time-lapse recording shown in
panels A-F. SO cells are identified by YFP fluorescence (green). External cells are identified by Ttk69immunostaining (red) and the sheath cell by Pros (blue). Note that the fragmented cell is Ttk69-positive (star in
F and G). (H, H') The anterior pI-daughter cell acquired a pI-cell fate. Lineage expressing Cdc25stg showing
anterior pI daughter division levelled for Pros (red in H and alone in H') and YFP (green in H). Note that the
division is oriented in the plane of the epithelium and was not associated with Pros expression (arrow), similar
to pI-mother cell division.
Fig. S6. Forcing premature pI-mitosis after expressing of Cdk1F did not accelerate the developmental
timing of neural-associated Pros-expression. (A) Percentage of sensory organs presenting 1 (blue), 2 (red), 3
(green) or 4 (violet) sensory cells as a function of developmental time after overexpression of Cdk1(WT) or
Cdk1F using neur-Gal4 as a driver. Note that after overexpression of Cdk1F, sensory organs develop earlier (of
about 3h) than after overexpression of Cdk1(WT). (B) Percentage of sensory organs presenting a Pros-positive
sensory cells as a function of developmental time after overexpression of Cdk1(WT) or Cdk1F using neur-Gal4
as a driver. Note that the timing of generation of a Pros-expressing cells is similar after overexpression of
Cdk1(WT) or Cdk1F.
Development • Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Development 143: doi:10.1242/dev.134270: Supplementary information
Movie 2. In vivo imaging of a neur>Cdk1F, H2B::YFP pupae during 10 hour period beginning at 14 hours
APF. H2B-YFP is in inverted fluorescence. Anterior is on the right and the view is dorsal.
Development • Supplementary information
Movie 1. In vivo imaging of a neur>Cdk1WT, H2B::YFP pupae (control) during 8h period beginning at 16
hours after pupal formation (APF). H2B-YFP using inverted fluorescence. Anterior is to the left and the
perspective is dorsal. In all films, each frame was obtained by combining a z-stack (composed of optical
sections separated by 1um) acquired every 3 minutes.
Development 143: doi:10.1242/dev.134270: Supplementary information
Movie 4. In vivo imaging of a neur> cdc25string, H2B::RFP, pon::GFP pupae during 10 hour period beginning at
12 hours APF. H2B-RFP is shown in red, PON-GFP is in green. Anterior is on the top-left and the view is
dorsal.
Development • Supplementary information
Movie 3. In vivo imaging of a neur>cdc25string, H2B::YFP pupae during 10 hour period beginning at 14 hours
APF. H2B-YFP is in inverted fluorescence. Anterior is on the top and the view is dorsal.