Fig. 5: Knockdown of USP10 sensitizes lung and ovarian cancer cells harboring mutant- or null-p53 to cisplatin by colony formation assays.

a, b Knockdown of USP10 sensitizes three lung and two ovarian cancer cell lines to cisplatin by colony formation assays. Control and USP10 stable knockdown H157, H23, H1299, SKOV3, and ES-2 cells were treated with cisplatin at the various concentrations (H157, 2 μM; H23, 0.5 μM; H1299, 1 μM; SKOV3 and ES-2, 0.3 μM) for 7–14 days. Colony formation assay was performed as described in the Methods. Colonies were visualized by crystal violet staining (a). Colonies numbers were quantified with OpenCFU software. The percentage of colony formation of indicated cells are shown in (b). c, d Inducible knockdown USP10 sensitizes H23 cells to cisplatin by colony formation assays. H23 control and stable inducible USP10 knockdown pool cells were plated at 100 cells/well into six-well dishes and cultured with or without Dox and then treated with cisplatin as indicated for ~14 days. Colonies were fixed, stained with methanol/crystal violet dye, and images captured (c). Colony numbers were counted by OpenCFU software (d). e, f Inhibition of USP10 by spautin 1 sensitizes H1299, H157 and H23 cells to cisplatin in colony formation assays. H1299, H157, and H23 cells were treated with various concentrations of cisplatin (0.5 μM for H1299 and H157; 0.1 μM for H23), USP10 inhibitor spautin-1 (1 μM for H1299, 2 μM for H157, and 0.5 μM for H23), or a combination of cisplatin and spautin-1 for 7–14 days. Colonies were fixed, stained with methanol/crystal violet dye, and images captured (e). The percentages of colony formation of the indicated cell lines are shown in (f). g, h USP10 inhibitor P22077 sensitizes H1299 cells to cisplatin in colony formation assays. Colony formation assays for H1299 were performed as e, f, except that spautin-1 was replaced by P22077 (2 μM) and that cisplatin concentration was 0.5 μM. For all the panels, each assay was performed in triplicate. The error bar represents standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001.