Fig. 3
RSL1D1 Promotes p53 Ubiquitination by Upregulating HDM2. RSL1D1-downregulated or -overexpressed HCT116p53+/+ cells was treated with MG-132 (25 μM) for 6 h to inhibit the degradation of ubiquitinated p53. Monoclonal antibody against p53 (DO-1) was used for IP and western blot analyses of ubiquitinated and non-ubiquitinated p53. A The levels of ubiquitinated p53 proteins were evaluated in siRSL1D1- or siNC-transfected HCT116p53+/+ cells. The levels of RSL1D1, p53, and HDM2 proteins in the input cell lysate were determined by western blot analysis and β-actin was used as a loading control. B The levels of ubiquitinated p53 proteins were evaluated in RSL1D1- or EGFP-overexpressed HCT116p53+/+ cells. The levels of RSL1D1, p53, and HDM2 proteins in the input cell lysate were determined by western blot analysis and β-actin was used as a loading control. C The levels of ubiquitinated p53 proteins were evaluated in RSL1D1-overexpressed HCT116p53+/+ cells transfected with siHDM2 or siNC. The protein levels of RSL1D1, p53, and HDM2 in the input cell lysate were determined by western blot analysis and β-actin was used as a loading control