Supplementary Figure 4: Effects of statins on oncogenic properties of human cancer cells.
From: DNAJA1 controls the fate of misfolded mutant p53 through the mevalonate pathway
(a) MTT assays using SK-Br-3, HT29 cells, as well as HCT116 sub-cell lines (p53wt, p53null, p53null+R175H, p53null+R273H) treated with different concentrations of DMSO or lovastatin. Error bars, means ± s.d. (n = 3 independent experiments), ∗P < 0.05, ∗∗P < 0.01, NS: not significant; Student’s t test (two-tailed). Results of WB for p53 in HCT116 p53wt, p53null+R175H, and p53null+R273H cells treated with DMSO or lovastatin are also shown on the right. (b) Colony formation assays using SK-Br-3, HT29, and HCT116 sub-cell lines seeded onto 6-well plates (500 cells) and treated with DMSO or lovastatin (4 μM) every other day for 10 days. Formed colonies were stained with crystal violet and counted. Representative images (left) and a summarized graph (right). Error bars, means ± s.d. (n = 3 independent experiments), ∗∗P < 0.01, NS: not significant; Student’s t test (two-tailed). (c) Representative images of PI-staining and flow cytometry using U2OS and KHOS/NP cells treated with MDSO (D) or lovastatin (L) for 72 h (left). Summarized graph showing percentages (%) of sub-G0/G1 fraction and G1/S ratio (right). Error bars, means ± s.d. (n = 3 independent experiments), ∗P < 0.05; Student’s t test (two-tailed). (d) Tumor formation assays in mice subcutaneously injected with CAL33 cells (1 × 106). When subcutaneous tumors reached 3 mm in diameter, mice were intraperitoneally injected with DMSO (D) or atorvastatin (A, 30 mg kg−1) daily, and tumors were measured every 2–3 day until day 21. Representative images of tumors are shown in insets. Error bars, means ± s.d. (n = 5 animals for each group), ∗P < 0.05; Student’s t test (two-tailed). Scale bar, 1 cm. (e) Body weights of mice bearing KHOS/NP-tumors treated with DMSO or atorvastatin (30 mg kg−1, daily). Error bars, means ± s.d. (n = 5 animals for each group). (f) Representative images of IHC for p53, Ki67, and cleaved caspase 3 using CAL33 tumors treated with DMSO or atorvastatin. Scale bar, 50 μm. Statistics source data for a–c are provided in Supplementary Table 1.
