Figure 2.

Hypertrophic stimulation enhances interaction of TFIIIB and pol III, but not TFIIIC, with class III genes in vivo. Cultured cardiomyocytes were treated with 10% FCS for 16 h to induce hypertrophic growth. Control cells were maintained in serum-free medium. ChIPs were performed with antibodies against TFIIIB (Brf1 and Bdp1), TFIIIC (110 and 220) and pol III (RPC155), as indicated. Negative control ChIPs used a TFIIB antibody and beads alone. (A) Association of each factor with 5S rRNA and tRNA^Leu genes, in unstimulated and hypertrophic cells, was analysed by PCR with gene-specific primers. Input genomic DNA (10 and 2% of that used in ChIPs) was analysed in parallel. (B) PCR products were quantified by densitometry and normalised to the appropriate input. The average fold increases in factor binding are represented (n=3; ^*significantly higher than control, P<0.05; ^**no significant difference from control).