Supplementary Figure 1: Screening strategy to identify statins as compounds that deplete p53^R175H.

(a) Saos2 (p53^null) cells were infected with a lentiviral vector encoding a chimeric fusion protein of p53^R175H and luciferase reporter (p53^R175H-Luc). Compounds that induce p53^R175H degradation are also expected to degrade the p53^R175H-Luc fusion protein, thereby reducing luciferase activities. The p53^R175H-Luc cells were exposed to chemical libraries containing ∼9,000 compounds (2.5 μM, bioactives and FDA approved drugs) and DMSO (control) for 24 h (1^st screening). Forty four (44) compounds showing reduced luciferase activity compared with DMSO were examined for the dose dependency in the luciferase activity using p53^R175H-Luc cells, as well as for a counter screening using Saos2 cells expressing only luciferase reporter (Luc, 2nd screening). Top 10 compounds that preferentially reduced luciferase activity in p53^R175H-Luc cells with dose dependency were selected. (b) Representative results of WB for p53 and vinculin using MG63 (p53^null) exogenously expressing p53^R175H or SK-Br-3 (p53^R175H) and U2OS (p53^wt) treated with 7 compounds other than statins for 24 h. (c) WB for p53 and vinculin using SK-Br-3 cells treated with DMSO (D) or lovastatin (L; 4 μM) for indicated time points (h: h). NT, not treated. (d) WB for p53 and vinculin using SJSA-1 (p53^wt) cells infected with retroviral vector encoding control empty (control) or p53^R175H cDNA (+ p53^R175H) and treated with D or L (4 μM) for 24 h. (e) WB for p53 and vinculin using H-2087 (p53^{V 157F}) cells treated with D or L for 24 h.