Figure 5

Overexpression of TAF3 decreases the level of p53 protein but not p53 mRNA. (A) U2OS cells were untransfected (U) or co-transfected with various amounts of plasmid expressing mTAF3 (0, 1, 2, 4 μg as indicated above the panel), and empty vector to a total of 4 μg DNA. p53 protein level in the cells was examined by Western blotting and the same blot was immunoblotted with anti-actin antibody for internal control. (B) HeLa cells were co-transfected with p53 expression construct, GFP-expression construct for control, and empty vector (0) or 4 μg of plasmid expressing mTAF3 (4). Cell lysates were prepared 24 h after transfection and analysed by Western blotting using anti-TAF3, anti-p53, or anti-GFP antibodies as indicated. (C) U2OS cells were co-transfected with various amounts of plasmid expressing mTAF3 (0, 1, 2, 4 μg) and empty vector to a total of 4 μg DNA. p53 and for internal control, GAPDH mRNA level was examined by QPCR, p53 mRNA level was normalized to GAPDH mRNA level in each sample, and expressed relative to normalized p53 mRNA level in vector-transfected control.