Fig. 4.

mSin3B directly represses E2F target genes transcription in vivo. (a) (Upper) PCR for mSin3B alleles on DNA from tail (before pIpC injection) or liver (10 days after pIpC injection) from Sin3B/Mx-Cre transgenic animals. (b) Western blot with the indicated antibodies from liver extracts 10 days after pIpC injection from mSin3B^L/+Mx-Cre⁺ mice (+/−) or mSin3B^L/−Mx-Cre⁺ mice (−/−). *, Nonspecific band. (c) qPCR analysis of mRNA abundance in mSin3B^+/− or mSin3B^−/− livers as generated in a. Shown is the average abundance of cDNAs after RT-PCR corresponding to the indicated transcripts, normalized to β-2-microglobulin. Three animals for each genotype were used. (d) ChIP assay on the promoter of the indicated E2F target genes and GAPDH, using liver extracts from mSin3B^L/+Mx-Cre⁺ mice (+/−) or mSin3B^L/−Mx-Cre⁺ mice (−/−) with control antibody (IgG, white bars) or anti-mSin3B antibody (black bars). Shown is the average of two independent experiments performed in duplicate. (e) Fold changes of the ChIP signal with E2F4 and HDAC1 antibodies on the promoter of cdc2a and brd8, in liver extracts from mSin3B^L/−;Mx-Cre⁺ mice (−/−) compared with liver extracts from mSin3B^L/+;Mx-Cre⁺ mice (+/−). The results are normalized to the corresponding H3 abundance. Shown is a representative experiment.