Fig. S1
siControl E2
G1: 45.7% S: 26.9%G2-M: 27.4%
siER E2
G1: 70.9% S: 9.9%G2-M: 19.2%
G1: 57.1% S: 12.0%G2-M: 30.9%
siRNF31 E2
A
B siRNF31 siControl siRNF31+ ERα plasmid
G1: 57.6% S: 22.6%G2-M: 20.3%
G1: 61.7% S: 13.8%G2-M: 24.9%
G1: 55.4% S: 18.9%G2-M: 26.2%
Fig. S2
siControl siRNF31
T47D
RNF31
ER
GAPDH
RNF31
ER
GAPDH
siControl siRNF31
MDAMB175
120 kDa
66 kDa
37 kDa
120 kDa
66 kDa
37 kDa
120 kDa
66 kDa
37 kDa
siCon
trol
siRNF31
SM
ARTpool
siRNF31
oligo
1
RNF31
ER
GAPDH
A
B
siRNF31
oligo
2
1.2
1.0
0.8
0.6
0.4
0.2
0
Rel
ativ
e m
RN
A le
vel
RNF31 (mRNA)
Fig. S3
A B
DC
NA
*** ***
Rel
ativ
e lu
cife
rase
act
ivity
NA
NA
Rel
ativ
e lu
cife
rase
act
ivity
Rel
ativ
e m
RN
A le
vel NA
NA NA
NA
Rel
ativ
e m
RN
A le
vel
***
***
***
***
Input
IP: RNF31
IB: ER
IP IgG IP RNF31
Input
IP IgG IP ER
IP: ER
IB: RNF31
Fig. S4
A BInput IgG IP RNF31
ER ER
IgG
Input IgG IP RNF31
Ig G
ER
Ig G
Input IgG IP RNF31
CInput IgG IP ERα Input IgG IP ERα Input IgG IP ERα
RNF31
Ig G
RNF31
Ig GIg G
RNF31
Fig. S5
nucleus cytosol nucleus cytosol nucleus cytosol nucleus cytosol
Fig. S6
RNF31 ER Merge Merge with nuclear stain
Fig. S7
Fol
d en
rich
men
t ove
r Ig
G c
ontr
ol
Gene name SequenceRNF31 Forward ACCCCCTATTGAGAGAGATTGCTRNF31 Reverse TGGAGCCTGGGACAGAGGERα Forward GCT ACG AAG TGG GAA TGA TGA AAGERα Reverse TCT GGC GCT TGT GTT TCA ACPS2 Forward CAT CGA CGT CCC TCC AGA AGA GPS2 Reverse CTC TGG GAC TAA TCA CCG TGC TGCyclin D1 Forward CAC GCG CAG ACC TTC GTCyclin D1 Reverse GGG CGG ATT GGA AAT GAA CADORA1 Forward GGA TCG ATA CCT CCG AGT CAADORA1 Reverse GAG AAT CCA GCA GCC AGC TA36B4 Forward GGCGACCTGGAAGTCCAACT36B4 Reverse CCATCAGCACCACAGCCTTCPKIB Forward CHIP TCA CTG CGG GAT ATT GGC TTA PKIB Reverse CHIP AGC TGG TCT TCT TCC TCC TAA ACT G ADORA1 Forward CHIP GAT GGA TGG GAA CAC ATT GGTADORA1 Reverse CHIP TGG TGG CGG AGC ACA AAERα Forward CHIP GGG ATC GCT CCA AAT CGAERα Reverse CHIP CTT GCC CTG ACA TTG GCT TAAPS2 Forward CHIP CCT CCC GCC AGG GTA AAT ACPS2 Reverse CHIP CCG GCC ATC TCT CAC TAT GAA18S Forward CHIP GCTTAATTTGACTCAACACGGGA18S Reverse CHIP AGCTATCAATCTGTCAATCCTGTC
Real-time PCR primers used in this study
Table S1
TCGA KMplot.com All samples All samples
Symbol r2 P-value Symbol r2 P-value
ACE 00.03 00.37 ACE 00.25 00.00ADA 00.03 00.31 ADA 00.06 00.01
ADORA1 00.04 00.19 ADORA1 00.11 00.00APOA1 00.17 00.00 APOA1 00.08 00.00AREG 00.07 00.05 AREG NA NA
AURKA 00.08 00.01 AURKA -0.09 01.00BTG2 00.06 00.08 BTG2 00.14 00.00CCNG2 00.02 00.53 CCNG2 -0.10 01.00CSF1R 00.01 00.05 CSF1R 00.21 00.00
CYP1A1 00.02 00.04 CYP1A1 00.22 00.00
ESR1 00.18 00.00 ESR1 00.13 00.00GJB2 00.00 00.06 GJB2 NA NA
GREB1 00.21 00.00 GREB1 00.11 00.00HMOX1 00.01 00.05 HMOX1 00.01 00.37IGFBP4 00.14 00.00 IGFBP4 00.20 00.00
KCNMA1 00.07 00.03 KCNMA1 00.07 00.00PDZK1 00.18 00.00 PDZK1 NA NASGK3 00.03 00.34 SGK3 NA NATFF1 00.24 00.00 TFF1 00.06 00.00
TGFB3 00.10 00.00 TGFB3 00.17 00.00
ACE2 -0.17 00.00 ACE2 -0.01 00.04ACPP -0.01 00.04 ACPP 00.18 00.00
CAV2 -0.30 00.00 CAV2 -0.08 01.00
CCNE1 -0.10 00.00 CCNE1 -0.09 01.00CD36 -0.16 00.00 CD36 -0.06 01.00
CDKN1A -0.02 00.57 CDKN1A 00.14 00.00CXCL1 -0.12 00.00 CXCL1 -0.02 00.05CXCL12 -0.09 00.01 CXCL12 00.00 00.45
CXCR4 -0.02 00.58 CXCR4 -0.10 01.00CXCR7 -0.17 00.00 CXCR7 -0.03 00.06CYP19A1 -0.07 00.02 CYP19A1 00.16 00.00CYP1B1 -0.18 00.00 CYP1B1 -0.07 01.00
EGFR -0.21 00.00 EGFR 00.02 00.17
EGR3 -0.01 00.06 EGR3 00.10 00.00NR5A2 -0.14 00.00 NR5A2 00.17 00.00
PPARA -0.20 00.00 PPARA 00.13 00.00REL -0.07 00.02 REL -0.01 00.04S100A8 -0.15 00.00 S100A8 -0.10 01.00
TRPV6 -0.09 00.00 TRPV6 -0.03 00.06WISP2 -0.03 00.39 WISP2 00.06 00.01
Table S2RNF31 expression correlates with expression of known ERα target genes
Yellow: p-value < 0.05; Red: positive correlation; Green: negative correlation