Fig. 6.

CCN5 binds and inhibits the TGF-βRII promoter. (A) 5′-flanking region deletion mutants of the TGF-βRII promoter-luciferase constructs were transiently transfected into 293T cells with 0, 0.5, and 1 μg of plasmid expressing hCCN5 and 0.1 μg of RSV-β-galactosidase construct as an internal control. At 48 h posttransfection, cells were harvested and analyzed for luciferase and β-galactosidase activities. Luciferase was expressed as mean ± SD of triplicates from a representative experiment performed at least three times. Significant differences: *, P < 0.05 versus control. (B) Cross-linked sheared chromatin from the MCF-7-sh-scrambled cells or from the MCF-7-sh-CCN5 and MDA-MB-231 cells were immunoprecipitated with the indicated specific antibodies. DNA was analyzed by PCR using primers to amplify the hTGF-βRII promoter region. Results shown are representative of three independent experiments.