Fig. 5
RSL1D1 Colocalizes with p53 and HDM2 and Negatively Regulates p53 in HCT116p53+/+ Cells under Normal and Nucleolar Stress Conditions. A, B IFA was performed to investigate the sub-cellular localization of RSL1D1 (red) (A, B), p53 (green) (A), and HDM2 (green) (B) in HCT116p53+/+ cells treated or untreated with 5 nM Act-D for 24 h. C, D IFA was performed to investigate the sub-cellular localization of RSL1D1 (red) (C, D), p53 (green) (C), and HDM2 (green) (D) in the siRSL1D1- or siNC-transfected HCT116p53+/+ cells. A-D The nuclei were stained with Hoechst (blue). Yellow color indicates the colocalization of RSL1D1 either with p53 (A, C) or HDM2 (B, D). Scale bars: 5 μm. E The levels of RSL1D1, p53, and HDM2 proteins were measured by western blot analysis in HCT116p53+/+ cells transfected with siRSL1D1 or siNC. The cells were treated or untreated with 5 nM Act-D for 24 h. β-actin was set as a loading control