Figure 5
From: Def defines a conserved nucleolar pathway that leads p53 to proteasome-independent degradation
Def-mediated p53 degradation depends on cysteine protease, CAPN3. (A) Western blot of p53 and Def for comparing the effect of knockdown of CAPN3, -6, -7 and -8 by their specific siRNAs on hu-Def-mediated p53 degradation. (B) qPCR showing the efficiency of knockdown of capn6, capn7 and capn8 by their respective siRNAs. The relative expression level of the genes was shown in fold change as normalized against human elf1a. (C) Western blot of p53, Def, Hdm2 and CAPN3 in MCF-7 cells treated with CAPN3-siRNA. (D) Western blot of p53 and CAPN3 in CAPN3-siRNA treated MCF-7 and HepG2 cells at 24 hpt. (E) Western blot of p53, CAPN3 and Def in cells transfected with CAPN3, p53 overexpression plasmids and hu-Def siRNA. (F) Co-IP analysis of interaction between hu-Def and CAPN3 in H1299 cells. Cells were transfected with plasmids as indicated. In (A, and C-E), fold changes in expression for p53, hu-Def or CAPN3 against their respective controls (set as 1) are shown under their corresponding panels. GAPDH was used as the normalization control.
