Protein target / Antibody / Dilution / Company
BRCA1 / D9 / 1:100 / Santa Cruz Biotechnology
GFP / Jl-8 / 1:3000 / Clontech
Cleaved Caspase-3 / Asp175 antibody, 9661 / 1:400 / Cell Signaling Technology
Bcl2 / 2872 / 1:1000 / Cell Signaling Technology
XIAP / 1:1000 / Cell Signaling Technology
p65 / sc-372 / 1:1000 / Santa Cruz Biotechnology
p50 / sc-8414 / 1:1000 / Santa Cruz Biotechnology
GAPDH / 6G5 / 1:5000 / Sigma
Actin / AC-74 / 1:5000 / Sigma
b-tubulin / 1:2000 / Sigma
TF11D (TBP) / SC-204 / 1:1000 / Santa Cruz Biotechnology

Supplementary Table 1: Antibodies used for Western Blotting

Gene / Primers
BRCA1 / Forward 5’ ATCATTCACCCTTGGCACA
Reverse 5’ GGAAGCCATTGTCCTCTGTC
P65 / Forward 5’ ACCGCTGCATCCACAGTT
Reverse 5’ GGATGCGCTGACTGATAGC
Bcl2 / Forward 5’ GGATGACTGAGTACCTGAACC
Reverse 5’ GGCCGTACAGTTCCACAAAGG
XIAP / Forward 5’ GCAAGAGCTCAAGGAGACCA
Reverse 5’ AAGGGTATTAGGATGGGAGTTCA
p50 / Forward 5’ CTGGCAGCTCTTCTCAAAGC
Reverse 5’ TCCAGGTCATAGAGAGGCTCA
IkBa / Forward 5’ GCTGATGTCAATGCTCAGGA
Reverse 5’ ACACCAGGTCAGGATTTTGC
Actin / Forward 5’AGACCTGTACGCCAACACAG
Reverse 5’GGAGCAATGATCTTGATCTTCA

Supplementary Table 2: Sequences of real time PCR primers


Supplementary Table 3: Antibodies used for ChIP assay

Protein Target / Antibody / Company
RNA Polymerase II / CTD4H8 / Millipore
BRCA1 / Ab-1 / Calbiochem
p52 / sc-848 / Santa Cruz Biotechnology
p50 / sc-114 / Santa Cruz Biotechnology
RelA/p65 / sc-372 / Santa Cruz Biotechnology
RelB / sc-226 / Santa Cruz Biotechnology
c-Rel / sc-71 / Santa Cruz Biotechnology
Rabbit IgG / DAKO
Mouse IgG / DAKO


Supplementary Table 4: Sequence of ChIP promoter primers

Promoter site / Primer / Reference
Bcl2 κ1 / Forward 5’ TGGAGCGCTGAGTTCTGCAT
Reverse 5’ GACAGGTGACTCTGCACGTTTTA / (1)
Bcl2 P1 / Forward 5’ CAGAGAATGAAGTAAGAGGACAGGCAC
Reverse 5’ GCCTCCCGGAGCCCCGCCGCGCA / (2)
Bcl2 P2 / Forward 5’ GTGTACAGGGAAACGCACCT
Reverse 5’ TCTTTATTTCATGAGGCACGTTATT / (3)
Bcl2ns
(approx 2300 bp upstream of κ1) / Forward 5’ AGATAGAGTTTTGTCATGTTGTGC
Reverse 5’ GGTGGGTGTATTGCTTGAGG
XIAP / Forward 5’ TGCCTGCTTAAATATTACTTTCCTCAAAA
Reverse 5’ ACTACACGACCGCTAAGAAACATTCT / (4)
XIAPns
(approx 3000 bp upstream) / Forward 5’ TCAGGATAATTATAGACTTGGAAAGGT
Reverse 5’ CCTATCACTTAAAGGAACTTCAGCA

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4. Mohapatra S, Chu B, Wei S, et al. Roscovitine inhibits STAT5 activity and induces apoptosis in the human leukemia virus type 1-transformed cell line MT-2. Cancer Res 2003; 63: 8523-30.

Supplementary Table 5: Statistical significance of differences in IC50 values between HCC-EV and BR cells transfected with either Scr, p65 or p50 siRNA, and treated with etoposide or camptothecin.

Cell type and transfection / Etoposide
treatment / Camptothecin treatment
HCC-BR vs HCC-EV siScr / P<0.001 / P<0.001
HCC-BR vs HCC-BR sip65 / P<0.0013 / P<0.001
HCC-BR vs HCC-BR sip50 / P<0.0013 / P<0.001
HCC-EV vs HCC-EV sip65 / P=0.6371 / P=0.08
HCC-EV vs HCC-EV sip50 / P=0.768 / P=0.226

Survival was assayed by colony count assay, associated survival curves are shown in Figure 7C. P values were calculated as described in Materials and Methods.