Figure 1.

Suv39h1 inhibits MyoD-mediated transcription in an HMT activity dependent manner. (A) C3H10T1/2 cells were cotransfected with indicated reporters and vectors encoding MyoD and Suv39h1. (B) C3H10T1/2 cells were cotransfected with indicated reporter, vectors encoding MyoD (0.2 μg) and an increased concentration of Suv39h1 (0.4, 0.6 and 0.8 μg). (C) C2C12 cells were cotransfected with the indicated reporters and Suv39h1 expression vector. (D, E) C3H10T1/2 or C2C12 cells were cotransfected with indicated reporters plus vector encoding enzymatic active (wt) or inactive H324K version of Suv39h1 in the presence of vector expressing MyoD. (F) Chromatin integrated control (C2-Em-LacZ) or 4RE-LacZ (C2-4RE-LacZ) cells were transfeted with active (wt) and inactive H324K version of Suv39h1. Normalized reporter activity was expressed as fold activation. (G) 293T cells were transfected with equal amount of plasmid encoding Flag-tagged wt or mutant version (H324K) of Suv39h1. Their expression levels were monitored by immunoblot analysis of equal amount of cell extracts using Flag antibodies.