Supplementary Figure 3: Both MVP and MVA-5PP rescue cholesterol production that is inhibited by statins, whereas only MVP, but not MVA-5PP, nullifies statin-induced mutp53 degradation.

(a) WB for p53 and vinculin using KHOS/NP cells treated with DMSO (D) or lovastatin (L; 4 μM) with or without supplementation with MVA-5PP (100 and 200 μM). (b) WB for p53 and vinculin using SK-Br-3 (p53^R175H) cells treated with cyclohexamide (CHX, 50 nM) for indicated time period, following pre-incubation with D, L (4 μM), L + MVP (200 μM), or L + MVA-5PP (200 μM) for 12 h (left). In Supplementary Fig. 2f and 3b, the same samples for DMSO and lovastatin treatments were used till 8 h-time points, and hence, the same data are used in both figures. Graph showing relative p53^R175H levels with time compared to those without CHX treatment (right). Error bars, means ± s.d. (n = 3 independent experiments). (c) Measurement of intracellular cholesterol. SK-Br-3 (left) and KHOS/NP (right) cells were treated with L, along with MVP (200 μM) or MVA-5PP (200 μM) for 24 h, and the levels of cholesterol were determined using the Total Cholesterol Assay Kit (Colorimetric; Cell Biolabs, San Diego, CA). Graphs showing cholesterol values relative to that in D-treated cells. Error bars, means ± s.d. (n = 3 independent experiments), ^∗P < 0.05, ^∗∗P < 0.01; Student’s t test (two-tailed). (d) Measurement of intracellular cholesterol levels in CAL33 cells infected with non-silencing control or MVK shRNA-encoding lentiviral vectors and treated with vehicle (water), MVA (200 μM), or MVP (200 μM). Error bars, means ± s.d. (n = 3 independent experiments), ^∗P < 0.05, ^∗∗P < 0.01, NS: not significant; Student’s t test (two-tailed). Statistics source data for b, c, and d are provided in Supplementary Table 1.