Supplementary data

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					                                  Supplementary data




Supp. Fig. 1:

A: w/t and p52/100 null MEF cells were fractionated by centrifugal elutriation and

stained with propidium iodide for analysis by FACS. A representative distribution of

cells at different cell cycle stages is shown.

B: Western blot showing NF-kB subunit levels in wild type, nfkb2 (p52/p100) -/- and

rela -/- MEFs.

C: Quantitative RT-PCR analysis was performed using primers specific to Cyclin D1,

Skp2, c-Myc and GAPDH control, using total RNA prepared from wild type and p52

null MEF cells following centrifugal elutriation.

D: U2OS cells were fractionated by centrifugal elutriation and stained with propidium

iodide and with antibodies to total p100, S866 phosphorylated p100 or an IgG control

as indicated, for analysis by FACS.

E: Semi-quantitative PCR analysis of Bcl-xL, Bcl-2, A20 and Bcl-3 mRNA levels in

fractions from elutriated U2OS cells.

F: U2OS cells were transfected with an siRNA targeting Bcl-3. mRNA expression

were analyzed as shown.



Supp. Fig. 2.

A & B: Analysis of Cyclin D1, Skp2, c-Myc and p52/p100 protein and mRNA

expression in U2OS (passage 13 and 26), MCF7 and HUT78 cells.

C: Table summarizing FACS analysis data showing the cell cycle distribution of

Cyclin D1, c-Myc and Cyclin B1 protein in U2OS (passage 13 and 26), MCF7 and

HUT78 cells. Cells were stained with antibodies to the indicated proteins and with

propidium iodide for DNA content. Numbers shown are the percentage of cells

scored by propidium iodide staining as being at a particular cell cycle stage that also
showed staining for the target protein.


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D: U2OS (passage 14 and 26), MCF7 and HUT78 cells were separated by elutriation

and fractions were analysed by western blotting for the indicated proteins.




Supp. Fig. 3:

Analysis of NF-B subunit, coactivator and repressor recruitment to the promoters of

NF-B target genes in U2OS cells. ChIP analysis of the Cyclin D1, Skp2, c-Myc and

GAPDH promoters was performed by Quantitative PCR using antibodies to the

indicated protein, in U2OS cells following centrifugal elutriation.



Supp. Fig. 4:

A: w/t and RelA null MEF cells were fractionated by centrifugal elutriation and

stained with propidium iodide for analysis by FACS. A representative distribution of

cells at different cell cycle stages is shown.

B: Analysis of NF-B subunit, coactivator and repressor recruitment to the promoters

of NF-B target genes in wild type, nfkb2 null and RelA null MEF cells following

centrifugal elutriation. ChIP analysis of the Cyclin D1, Skp2, c-Myc and GAPDH

promoters was performed by Quantitative PCR using antibodies to the indicated

protein.



Supp. Fig. 5:

A: Western blot analysis of whole cell MEF extracts following centrifugal elutriation

for S468 and S536 phosphorylated RelA.

B: IKK or IKK were immunoprecipitated from whole cell lysates prepared from

elutriated U2OS cells. The immunoprecipitate was incubated with whole cell lysate

from serum starved U2OS cells.          These lysates were then analysed by western

blotting with antibodies to S866 phosphorylated p100 and S468 or S536

phosphorylated RelA as indicated




                                             2
C: U2OS Nuclear, cytoplasmic and whole cells protein extracts following centrifugal

elutriation were pooled according to the cell cycle stage and subjected to western blot

analysis using antibodies to Chk1.



Supp. Fig. 6.

Control quantitative PCR analysis showing ATR, ATM, Chk1, Chk2 and IKK

mRNA levels following treatment of U2OS cells with the indicated siRNAs.



Supp. Fig. 7.

Chk1 inhibition of IKK activity requires Chk1 kianse activity. Experiment was

performed as in Figure 8B, except that either the Chk1 inhibitor Go6976 was included

(A) or purified Chk1 was inactivated by heat treatment (B). An additional control, the

inclusion of a non-hydrolysable ATP analogue, is also shown.



Supp. Fig. 8.

A: Analysis of NF-B subunit, HDAC1 and CBP recruitment to the promoters of NF-

B target genes in each cell cycle stage following Chk1 and Akt pathway inhibition.

ChIP analysis of the Cyclin D1, Skp2, c-Myc and GAPDH promoters, was performed

in U2OS following centrifugal elutriation of U2OS cells with or without Gö6976 or

LY294002 treatment.

B: RNA extracts were prepared from elutriated U2OS cells treated with Gö6976 or

LY294002 and subjected to semi-quantitative PCR analysis as indicated.



Supp. Fig. 9.

Cell cycle analysis of all elutriated U2OS cell fractions used in each Figure.



Supp. Fig. 10. Summary of the primers used for ChIP assays in U2OS cells.




                                           3
Schematic diagram of the human Cyclin D1 (A), c-Myc (B), Skp2 (C), Bcl-2 (D) and

Bcl-xL (E) promoters. The relative positions of previously described NF-B binding

sites () are shown.

A: 3 NF-B binding sites (-858, -749 and -39) have been described on the Cyclin D1

promoter (Guttridge et al, 1999; Joyce et al, 1999; Westerheide et al, 2001).

B: Two NF-B binding sites (-1095 and +449) have been described on the human c-

Myc gene (Duyao et al, 1990; Kessler et al, 1992).

C: The NF-B binding sites on Skp2 promoter have not been characterized but it was

previously shown that ChIP primers over the transcription initiation site can detect

NF-B subunit binding (Schneider et al, 2006) and these were used in this analysis.

D: Three NF-B binding sites have been described on Bcl2 gene, one in the upstream

promoter (-2445) (P1) and the others are in the P2 region (-212 and -174) (Catz &

Johnson, 2001; Xiang et al, 2006).

E: The primers used to the 1 NF-B binding site on the Bcl-xL promoter (Chen et al,

2000) have been described previously (Campbell et al, 2004)

F: Three NF-kB binding sites have been described on the IkBa promoter (-304, -50

and -19) (Ito et al, 1994).




                                           4
Supplementary Information: Materials and methods



Inhibitors

IKK inhibitors BAY 11-7082 (10M) and IKKi IV ([5-(p-Fluorophenyl)-2-

ureido]thiophene-3-carboxamide), together with the Chk1 inhibitor Go6976 were

purchased from Merck Biosciences.     Caffeine and aphidicolin were purchased from

Sigma Aldrich. PI-3 kinase inhibitor LY 294002 was purchased from Promega.

Concentrations used were: Go6796: 1μM on the cells and 0.1μM for the kinase

assay; LY294002: 50μM; Bay 11-7082: 10μM; IKKi IV: 50nM; caffeine:1mM;

aphidicolin: 2M.



Western blot

Western blots were performed essentially as described previously (Rocha et al, 2003).

For assays involving detection of phosphorylated proteins, 50-150g of protein

extracts were loaded per well.



Flow cytometric analysis of cell cycle distribution and proteins.

Adherent and detached cells were harvested, pooled, washed once in phosphate-

buffered saline (PBS), and fixed by incubation in 1% paraformaldehyde/PBS (pH 6.8)

for 10 min. Cells were washed once in PBS 0.1% tween-20 and then permeabilised in

70% ethanol (vol/vol). Cells were then washed twice in PBS (plus 0.1% (vol/vol)

tween-20) and resuspended in PBS containing 0.1% (vol/vol) tween-20 and 1% BSA

(blocking buffer). After incubation at room temperature for 15 min, cells were

incubated with blocking buffer containing antibody for 20 min. The dilution of the

antibodies used was 1:200 for anti-p52, anti-phospho-serine p100, anti-Cyclin D1,

anti-Cyclin B1 and anti-c-Myc antibodies. All secondary antibodies (labelled with

FITC or Cy-5) were purchased from Jackson Immunoresearch and used at 1:200

dilution. The cells were resuspended in PBS containing 0.1% (vol/vol) tween-20, 50
µg/ml of propidium iodide and 50 µg/ml of RNase A for 20 min at room temperature,


                                         5
and analyzed for cell cycle distribution with a FACS Calibur flow cytometer and

CellQuest software (Becton Dickinson). For all antibodies used in this study,

appropriate controls were performed. Cells were either stained with primary

antibodies without secondary antibodies to control for auto-fluorescence or stained

with secondary antibodies alone to control for background staining.



Chromatin Immunoprecipitation (ChIP)

Cells, either grown to 70% confluency or harvested and subjected to elutriation, were

cross-linked with 1% formaldehyde at room temperature for 10 min. Cells were

washed twice with 10 ml of ice-cold phosphate-buffered saline and then scraped into

0.5 ml of lysis buffer (1% SDS, 10 mM EDTA, 50 mM Tris-HCl, pH 8.1, 1 mM

PMSF, 1 g/ml leupeptin, 1 g/ml aprotinin) and left on ice for 10 minutes. Samples

were then sonicated at 4°C seven times. Each sonication was for 20 seconds with a

40 seconds gap between each sonication.           Supernatants were recovered by

centrifugation at 12,000 rpm in an eppendorf microfuge for 10 min at 4°C before

being diluted 4-8 fold in dilution buffer (1% Triton X-100, 2 mM EDTA, 150 mM

NaCl, 20 mM Tris-HCl, pH 8.1). Samples were then pre-cleared for 2 hours at 4°C

with 2 g of sheared salmon sperm DNA and 20 l of protein A or G-Sepharose

(50% slurry). At this stage, 20l of the material was kept as Input material.

Immunoprecipitations were performed overnight with specific antibodies (1g), with

the addition of NP-40 detergent to a final concentration of 0.5%. The immune

complexes were captured by incubation with 20 l of protein A or G-Sepharose (50%

slurry) and 2 g salmon sperm DNA for 1 hour at 4°C. The immunoprecipitates were

washed sequentially for 5 minutes each at 4°C in TSE 1 (0.1% SDS, 1% Triton X-

100, 2 mM EDTA, 20 mM Tris-HCl, pH 8.1, 150 mM NaCl), TSE 2 (0.1% SDS, 1%

Triton X-100, 2 mM EDTA, 20 mM Tris-HCl, pH 8.1, 500 mM NaCl), and Buffer 3

(0.25 M LiCl, 1% Nonidet P-40, 1% deoxycholate, 1 mM EDTA, 10 mM Tris-HCl,

pH 8.1). Beads were washed twice with TE buffer (10mM Tris-HCl, 1mM EDTA)
and eluted with 500 l of Elution Buffer (1% SDS, 0.1 M NaHCO3). For ReChIP


                                          6
experiments 25µl of ReChIP buffer (Dilution Buffer, 10mM DTT) was added to

beads following washes and incubated at 37°C for 50 minutes. The sample was then

diluted 40 times in dilution buffer and immunoprecipitations, washes and elution were

performed as before.

       To reverse the crosslinks, samples, including 'Input', were incubated at 65 °C

overnight. DNA was precipitated using classical Phenol-Chloroform procedures. For

PCR, 5l of DNA was used from an 80µl DNA preparation and subjected to 30

cycles of PCR amplifications. Control regions for GAPDH gene were subjected to 35

cycles of PCR Amplification.



Quantitative PCR analysis

Total RNA was extracted with the Nucleospin RNA II isolation system (Macherey

Nagel; 740955), according to the manufacturer’s directions. For reverse transcriptase

PCR (RT-PCR), 1g RNA sample were reverse transcribed with Quantitect Reverse

Transcription Kit (QIAgen; 205313). The cDNA stock was diluted by 200 and 5l

was used for PCR with GoTaq flexi DNA polymerase (Promega; M8305).

Quantitative PCR data was generated on a Rotor-Gene 3000 (Corbett Research) using

the following experimental settings: Hold 50°C for 3 min; Hold 95°C 10 min; Cycling

(95°C for 20 sec; 55°C for 20 sec; 72°C for 20 sec with fluorescence measurement) ×

45; Melting Curve 50–99°C with a heating rate of 1°C every 5 sec. All values were

calculated relative to untreated levels and normalised to GAPDH levels using the

Pfaffl method (Pfaffl, 2001). Each RNA sample was assayed in triplicate and the

results shown are representative of three separate experiments.



Kinase assay

Cells were elutriated as indicated previously. Pellets were lysed in lysis buffer (25

mM Hepes (pH 7.9), 300 mM KCl, 3 mM EDTA, 0.2 mM EDTA, 10% glycerol

(vol/vol), 0.5% NP-40, 0.5 mM PMSF, 1 mM sodium orthovanadate, 1 µg of
leupeptin/ml, 1 µg of aprotinin/ml, 10 mM p-nitrophenyl phosphate, 10 mM sodium


                                          7
fluoride). Supernatants were diluted further in lysis buffer and precleared with

Protein G-sepharose for 1hr. Precleared extracts were immunoprecipitated with 1 µg

of antibody overnight at 4°C. Protein G-Sepharose beads (Amersham Pharmacia

Biotech) were then added to each tube, and the samples were rotated for 60 min at

4°C. Afterward, samples were washed twice in lysis buffer, once in TE buffer (10mM

Tris-HCl, 1mM EDTA) and once in kinase buffer (50 mM HEPES (pH 7.7), 20 mM

MgCl2, 200 mM NaCl2, 10% glycerol, 10 µM ATP). The kinase activity was assayed

in kinase buffer by incubating with 25 µg whole cell extracts from serum starved cells

for 15 min at 30°C, supplemented with purified recombinant GST-Chk1 if required

(provided by Dr John Rouse, Dundee).



Antibodies

Antibodies used were: anti-p21 (sc-397, Santa Cruz), anti-p52 monoclonal and anti-

p52 polyclonal (05-361 & 06-413, Upstate Biotechnology), anti-p52 polyclonal (sc-

848, Santa Cruz), anti-p50 NF-B (sc-114, Santa Cruz), anti-RelA (sc-372, Santa

Cruz), anti-RelB (sc-226, Santa Cruz), anti-c-Rel (sc-71, Santa Cruz), anti-

IKK(sc-7607, Santa Cruz), anti--actin (A5441, Sigma), anti-Cyclin D1 (556470,

BD Pharmingen), anti-Skp2 (sc-7164, Santa Cruz), anti-c-Myc (sc-42, sc764, sc789;

Santa Cruz), anti-p21WAF1 (PC55-100ug, Oncogene Research Product), anti-Cyclin

A (CC02-20UG, Oncogene research product), anti-Cyclin B1 (sc752, Santa Cruz;

V152, Corning), anti-Chk1 (sc-7898, Santa Cruz), anti-Akt (sc-8312, Santa Cruz),

Cyclin E (CC05-20UG, Oncogene Research Product), CDC25C (sc-327, Santa Cruz)

Antibodies used in ChIP assays were: anti-p52 polyclonal (sc-848, Santa Cruz), anti-

p50 NF-B (sc-114, Santa Cruz), anti-RelA (sc-372, Santa Cruz), anti-RelB (sc-226,

Santa Cruz), anti-c-Rel (sc-71, Santa Cruz), anti-Gal4 (sc-510, Santa Cruz), anti-CBP

(sc-369, Santa Cruz), anti-Bcl-3 (sc-185, Santa Cruz), anti-mSin3A (sc-767, Santa

Cruz), and anti-HDAC1 (sc-6298, Santa Cruz).

Phospho-antibodies used in ChIP assays, western blot and co-immunoprecipitation
were   S866-p100    (4881S,    Cell   Signaling;   ab31474-100,   Abcam),     S180/1-


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IKK(2681, Cell Signaling), S280-Chk1 (2347S, Cell Signaling), S317-Chk1

(2344S, Cell Signaling), S345-Chk1 (2341S, Cell Signaling), T308-Akt (4056, Cell

Signaling), S473-Akt (4058, Cell Signaling), S468-RelA (3039L, Cell Signaling),

S536-RelA (3031S, Cell Signaling). The Goat T505-RelA phospho-specific antibody

was raised in collaboration with Active Motif and has been described previously

(Campbell et al, 2006).       The rabbit T505-RelA antibody has been described

previously (Rocha et al, 2005).



Oligonucleotides

ChIP:

Cyclin D1-human:

quantitative pcr primer set

FOR-AGTCCGTGTGACGTTACTGTTGT

REV-CTCCCGCTCCCATTCTCT

semi quantitative pcr primer set

FOR-ATTCTCTGCCGGCTTTGATC

REV-CGCTCGGCTCTCGCTTCT

Cyclin D1-Murine: FOR-CTCTGCTACTGCGCCGACA

REV-CACACGGACTACAGGGGAGTTTT

Skp2-Human: FOR-ACGAAGCGGGACGGAAACTA

REV-AGCTGCTCGCCTCCCAGAT

Skp2-Murine: FOR-CGTAGTCCTGCCTGGGTTCTT

REV-CGCAGGGAGTTGTGGGTAT

c-Myc-Human: FOR-ACTTTGCACTGGAACTTACAACAC

REV-CGAAAAAAATCCAGCGTCTAAG

c-Myc-Murine: FOR-GTCCTGGGAGGAAGGGGTTAA

REV-AGACGCGAGAATATGCCATGA

GAPDH-Human: FOR- CGGTGCGTGCCCAGTTG
REV- GCGACGCAAAAGAAGATG


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GAPDH-Murine: FOR-GCTACACTGAGGACCAGGTTG

REV-GCCCCTCCTGTTATTATGGGG

IkB-Human: FOR-CTTAGAAGTCTGGGGAAAGCAAAT

REV-GTAATCCTGTCCCTCTGCAAGT

BCL2-Human: FOR-TGGAGCGCTGAGTTCTGCAT

REV-GACAGGTGACTCTGCACCGTTTTA

Bcl-xL-Human: FOR-CCTCTCCCGACCTGTGATAC

REV-CCCCCGTCTTCTCCGAAATG

BCL-2: FOR-TGGAGCGCTGAGTTCTGCAT

REV-GACAGGTGACTCTGCACGTTTTA



RT-PCR:

Cyclin D1-Human: FOR-GTGCTGCGAAGTGGAAACC

REV-ATCCAGGTGGCGACGATCT

Cyclin D1-Murine: FOR-CAGAAGTGCGAAGAGGAGGTC

REV-TCATCTTAGAGGCCACGAACAT

Cyclin E-Human: FOR-TGAAGTGGTTAAGAAAGCCTCAG

REV-GAAGAAATCATGCACAGCATAGC

Cyclin A-Human: FOR-ACATTACAGATGATACCTACACCAAG

REV-CTTTGTCCCGTGACTGTGTAGAGTG

Skp2-Human: FOR-ATTGTCCGCAGGCCTAAGCTA

REV-TGCCATAGAGACTCATCAGACGC

Skp2-Murine: FOR-ATGGACTGCTCTCAAACCTCG

REV-CCTGGAAAGTTCTCCCGACTAA

c-Myc-Human: FOR-CCACAGCAAACCTCCTCACAG

REV-GCAGGATAGTCCTTCCGAGTG

c-Myc-Murine: FOR-TCTCCATCCTATGTTGCGGTC

REV-TCCAAGTAACTCGGTCATCATCT
p21WAF1-Human: FOR-CTGCCGCCGCCTCTTC


                             10
REV-CTGAGCGAGGCACAAGGGTA

ATR-Human: FOR-ACATTCCCTGATCCTACATCATG

REV-TTCAATAGATAACGGCAGTCCTG

ATM-Human: FOR-CCCCTTGTGTATGAGCAGGTG

REV-CGGATTATCCTGAGAAGCTC

Chk1-Human: FOR-ATCGATTCTGCTCCTCTAGC

REV-CATGTGGGCTGGGAAAAGCTG

Chk2-Human: FOR-CAAGAACCTGAGGACCAAGA

REV-CAAAGGTTCCATTGCCACTG

IKK-Human: FOR-GTTTGCAAGCAGAAGGCGCTG

REV-GTCTAGGGCCGTGAAACTCTG
p50/105-Human: FOR-TCCCATGGTGGACTACCTGG
REV-ATAGGCAAGGTCAGGGTGC
p52/100-Human: FOR-AAGGACATGAGTGCCCAATTTAAC
REV-ATCATGGATGGGCTGGGAGG
RelA-Human: FOR-CTGCCGGGATGGCTTCTAT
REV-CCGCTTCTTCACACACTGGAT
RelB-Human: FOR-CATCGAGCTCCGGGATTGT
REV-CTTCAGGGACCCAGCGTTGTA
c-Rel-Human: FOR-AGAGGGGAATGCGTTTTAGATACA
REV-CAGGGAGAAAAACTTGAAAACACA
Bcl-XL-Human: FOR-GGTCGCATTGTGGCCTTTTTC
REV-TGCTGCATTGTTCCCATAGAG
Bcl2-Human: FOR-CGACGACTTCTCCCGCCGCTACCGC
REV-CCGCATGCTGGGGCCGTACAGTTCC
Bcl3-Human: FOR-GCAGATCTTGGACTCATGAGG
REV-CTGGGGTCAGAGTCCTGGGAG
A20-Human: FOR-CACGAGCCCGAGACTGATGAGG
REV-CTTCCCCTTGCTCGTCACTG
CDC25C-Human: FOR-GTATCTGGGAGGACACATCCAGGG
REV-CAAGTTGGTAGCCTGTTGGTTTG
GAPDH-Human: FOR-GGTCGTATTGGGCGCCTGGTCACC


                            11
REV-CACACCCATGACGAACATGGGGGC
GAPDH-Murine : FOR-GCTACACTGAGGACCAGGTTG
REV-GCCCCTCCTGTTATTATGGGG


                                      References

Campbell KJ, Rocha S, Perkins ND (2004) Active repression of antiapoptotic gene
expression by RelA(p65) NF- B. Mol Cell 13: 853-865

Campbell KJ, Witty JM, Rocha S, Perkins ND (2006) Cisplatin mimics ARF tumor
suppressor regulation of RelA (p65) nuclear factor-B transactivation. Cancer Res
66: 929-935

Catz SD, Johnson JL (2001) Transcriptional regulation of bcl-2 by nuclear factor B
and its significance in prostate cancer. Oncogene 20: 7342-7351

Chen C, Edelstein LC, Gelinas C (2000) The Rel/NF-B family directly activates
expression of the apoptosis inhibitor Bcl-x(L). Mol Cell Biol 20: 2687-2695

Duyao MP, Buckler AJ, Sonenshein GE (1990) Interaction of an NF-B-like factor
with a site upstream of the c-myc promoter. Proc Natl Acad Sci U S A 87: 4727-4731

Grdina DJ, Meistrich ML, Meyn RE, Johnson TS, White RA (1984) Cell synchrony
techniques. I. A comparison of methods. Cell Tissue Kinet 17: 223-236

Guttridge DC, Albanese C, Reuther JY, Pestell RG, Baldwin AS, Jr. (1999) NF-B
controls cell growth and differentiation through transcriptional regulation of cyclin
D1. Mol Cell Biol 19: 5785-5799

Ito CY, Kazantsev AG, Baldwin AS, Jr. (1994) Three NF- B sites in the IB-
promoter are required for induction of gene expression by TNF. Nucleic Acids Res
22: 3787-3792

Joyce D, Bouzahzah B, Fu M, Albanese C, D'Amico M, Steer J, Klein JU, Lee RJ,
Segall JE, Westwick JK, Der CJ, Pestell RG (1999) Integration of Rac-dependent
regulation of cyclin D1 transcription through a nuclear factor-B-dependent pathway.
J Biol Chem 274: 25245-25249

Kessler DJ, Spicer DB, La Rosa FA, Sonenshein GE (1992) A novel NF-B element
within exon 1 of the murine c-myc gene. Oncogene 7: 2447-2453

Pfaffl MW (2001) A new mathematical model for relative quantification in real-time
RT-PCR. Nucleic Acids Res 29: e45

Rocha S, Garrett MD, Campbell KJ, Schumm K, Perkins ND (2005) Regulation of
NF-B and p53 through activation of ATR and Chk1 by the ARF tumour suppressor.
Embo J 24: 1157-1169


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Rocha S, Martin AM, Meek DW, Perkins ND (2003) p53 represses cyclin D1
transcription through down regulation of Bcl-3 and inducing increased association of
the p52 NF-B subunit with histone deacetylase 1. Mol Cell Biol 23: 4713-4727

Schneider G, Saur D, Siveke JT, Fritsch R, Greten FR, Schmid RM (2006) IKK
controls p52/RelB at the skp2 gene promoter to regulate G1- to S-phase progression.
Embo J 25: 3801-3812

Westerheide SD, Mayo MW, Anest V, Hanson JL, Baldwin AS, Jr. (2001) The
putative oncoprotein Bcl-3 induces cyclin D1 to stimulate G(1) transition. Mol Cell
Biol 21: 8428-8436

White RA, Grdina DJ, Meistrich ML, Meyn RE, Johnson TS (1984) Cell synchrony
techniques. II. Analysis of cell progression data. Cell Tissue Kinet 17: 237-245

Xiang H, Wang J, Boxer LM (2006) Role of the cyclic AMP response element in the
bcl-2 promoter in the regulation of endogenous Bcl-2 expression and apoptosis in
murine B cells. Mol Cell Biol 26: 8599-8606




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