Figure 4.

TRIM45 stabilizes and activates p53 in glioma cells. (a) Lysates of TRIM45-overexpressing and control U87 and LN229 cells were immunoblotted with anti-p53, anti-PUMA, and anti-Flag antibodies. β-Actin was used as a loading control. (b) Lysates of TRIM45 KO and control U87 and LN229 cells were immunoblotted with anti-p53, anti-PUMA, and anti-TRIM45 antibodies. β-Actin was used as a loading control. (c and d) The mRNA levels of p53 target genes in TRIM45 overexpression and KO U87 cells were analyzed using real-time PCR. (e) Luciferase activity in U87 cells transfected with p53-luc, p53, and an empty vector or a vector expressing TRIM45 (0, 200, and 400 ng). Results are presented as reporter gene activity relative to Renilla luciferase activity. (f and g) Immunoblot analysis of extracts of TRIM45 KO or control U87 cells treated for various times (as indicated above each lane) with CHX. (h and i) Lysates of TRIM45 KO and control U87 cells treated with MG132 or dimethyl sulfoxide (DMSO) were immunoblotted with anti-p53 and anti-TRIM45 antibodies. β-Actin was used as a loading control. Each bar represents the mean±S.D. of three independent experiments. *P<0.05, **P<0.01, and ***P<0.001