Fig. 4: PDCD11 relies on HDM2 to ubiquitinate and degrade p53. | Oncogenesis

Fig. 4: PDCD11 relies on HDM2 to ubiquitinate and degrade p53.

From: Programmed cell death 11 modulates but not entirely relies on p53-HDM2 loop to facilitate G2/M transition in colorectal cancer cells

Fig. 4

A, B and D All the lentivirus-transduced cells were treated with Doxy to induce expression of shRNAs. C, E Cells were transfected with pBiFC-mCherryN159-FLAG or pBiFC-mCherryN159-FLAG-PDCD11. A Western blot analyses were performed to determine the levels of PDCD11 and p53 proteins in HCT116p53+/+ cells treated with 100 µg/mL CHX for indicated times. To facilitate comparison, p53 bands were obtained through a short exposure (SE) for shPDCD11 or long exposure (LE) for shLuc. GAPDH was used as a loading control. Relative quantification of the p53 levels was shown (n = 3). The values of the cells at 0 h were set to 1. B, C Ubiquitinated and non-ubiquitinated p53 was immunoprecipitated using anti-p53 (DO-1) from HCT116p53+/+ cells treated with MG-132 (20 µM, 8 h). Anti-Ubiquitin (P4D1) and anti-p53 (DO-1) were used to detect ubiquitinated and non-ubiquitinated p53, respectively. The levels of PDCD11/FLAG-PDCD11, p53, and HDM2 proteins in the input cell lysate were determined by western blotting and GAPDH was used as a loading control. D, E Western blot analyses were performed to determine the levels of PDCD11/FLAG-PDCD11 and p53 proteins in HCT116p53+/+ cells untreated or treated with Nutlin-3 (40 µM, 12 h). GAPDH was used as a loading control.

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