Table 1. List of genes with greater than 3-fold changes (in STAT3KO MEF) in response to IFN
Fold
change
Fold
change
(ST3KO/IFN
vs ST3KO)
(WT/IFN
vs WT)
VIPERIN
102.6
16.8
TYKI
44.9
8.1
USP18
MX2
IRGM2
G1P2
37.5
30.3
23.3
22.4
9.6
3.2
9.3
4.8
IGTP
18.7
8.6
IFI1
17.1
9.9
CXCL10
OCIL
14.7
10.5
9.2
2.7
TOR3A
10.3
8
STAT1
GBP4
IFIT3
IFI47
PRKR
ISGF3G
STAT2
9.2
8.7
8.6
7.4
7.3
6.7
6.5
4
3.7
3.5
2.8
6.7
5.6
4.3
OASL1
6.3
2.6
Gene name
Description
IFN-inducible antiviral protein
Thymidine kinase family LPSinducible member
ISG15-specific protease
Antiviral GTPase
IFN-inducible GTPase
ISG15
IFN -inducible GTP binding
protein
IFN-induced protein with
tetratricopeptide repeats 3
Chemokine
Osteoclast inhibitory lectin
ATP-dependant IFN response
protein
STAT
GTPase
IFN-inducible protein
IFN-inducible protein
Ser/Thr protein kinase
IFN regulatory factor
STAT
2’,5’-oligoadenylate
synthetase
ADAR
TREX1
BST2
IFI205
IRF1
ZC3HAV1
DAXX
PML
DDX58
6
2.2
5.5
4.7
4.1
4
4
3.8
3.5
3.2
3.3
2.3
4.3
2.9
2.8
2.5
3
2.4
adenosine deaminase, RNAspecific
DNase III
CD317, tetherin
IFN-inducible protein
IFN regulatory factor
Zinc finger antiviral protein
Death-associated protein
Ring finger protein, TRIM19
RIG-I
* RNA prepared from WT or STAT3KO MEFs that were treated without or with IFN 1000 U/ml for 2 h was
subjected to expression microarray analysis using Illumina MouseWG-6 v1.1 Expression BeadChip
The microarray data has been deposited to GEO accession # GSE25044
http://www.ncbi.nlm.nih.gov/gds/?term=GSE25044
Fig. S1
A.
WT (MEF)
IFND (h)
0 0.5 1
ST3KO (MEF)
3 6
9
0 0.5 1
3
6
9
pST1
ST1
pST2
ST2
pST3
ST3
Tubulin
B.
WT (BMM)
IFND(h)
0
1
4
6
ST3KO (BMM)
9
0
1
4
6
9
pST1
ST1
Tubulin
Fig. S1 Activation of STATs in WT and STAT3KO cells in response to IFN. WT
(left) or STAT3KO (right) MEFs (A) or BMMs (B) were treated with IFND 1000 U/ml
and 100 U/ml, respectively, for the indicated times. Total cell lysates were subjected to
immunoblotting using antibodies to pSTAT1, pSTAT2, pSTAT3, STAT1, STAT2,
STAT3 and tubulin for MEFs and pSTAT1, STAT1 and tubulin for BMMs.
Fig. S2
*
2
1
E.
6
5
4
0h
3
**
0
0h
5h
**
2
1
F.
WT
ST3KO
1
3
0
5h
TLR3
2
4
WT
ST3KO
3.5
3.0
2.5
0h
WT
ST3KO
2.0
1.5
0.0
**
0h
3
WT
ST3KO
2
**
1
0h
G.
MDA5
0.5
IRF1
4
0
5h
1.0
D.
5h
Relative mRNA (X10-2)
3
5
Relative mRNA (X10-2)
WT
ST3KO
6
iNOS
Relative mRNA (X10-2)
4
Relative mRNA (x10-3)
OAS
5
0
Relative mRNA (X10-4)
C.
B.
Relative mRNA (X10-4)
Relative mRNA (X10-2)
A.
8
6
4
**
2
0
0h
5h
h.p.i.
0 3 6 3 6
pST1
*
4
0
WT
ST3KO
IFNAR1 Ab
WT
ST3KO
6
2
8
IRF7
H.
RIG-I
12
10
5h
10
Tubulin
**
0h
pST2
5h
Fig. S2 Increased expression of IFN downstream genes in STAT3KO BMMs after EMCV infection. WT
(solid) or STAT3KO (empty) BMMs were infected with EMCV at an MOI of 10 for the indicated times. RNA
prepared from the cells was subjected to RT-QPCR using primers for OAS(A), iNOS(B), IRF1 (C), IRF7(D),
TLR3(E), MDA5(F), RIG-I(G) and E-actin. Relative mRNA was calculated by normalizing the values of specific
genes to that of E-actin. (H) STAT3KO (right) MEFs were pretreated without or with anti-IFNAR1 antibody 1
Pg/ml, followed by infection with EMCV at an MOI of 0.1 for the indicated times. Total cell lysates were then
subjected to immunoblotting using antibodies to pSTAT1, pSTAT2 and tubulin. h.p.i. hour post-infection