Figure 5.

PC1-CTT activation of RunX2 requires p300, and PC1-CTT enhances the TAZ-p300 association (A) HEK293 cells were transfected with either control shRNA (shControl) or shRNA directed against human p300 (shp300). mRNA knockdown efficiency was assessed by qRT-PCR. (B) HEK293 cells were transfected with shControl or shp300. After 48h, the cells were super-transfected with RunX2-luciferase and Renilla-luciferase alone, or in the presence of the PC1-CTT. Luciferase values were measured 24 h after RunX2-lucferase transfection (72 h total after initial shRNA treatment). (C) HEK293 cells were transfected with Myc-p300, FLAG-TAZ, and HA-PC1-CTT where indicated. Cell lysates were incubated with anti-FLAG beads, complexes were eluted in SDS-PAGE loading buffer and run on a 10% SDS- polyacrylamide gel and blotted with the indicated antibodies. Densitometry was performed on the immunoprecipitated p300 band using image analysis software (right panel). Results are expressed as mean ± SE from four independent experiments.