Fig. 4

OTUD3 facilitates tumorigenicity in human lung cancer cells. a OTUD3 was knocked down in lung cancer cell lines H1299 using shRNA. The protein levels of OTUD3 were analyzed by western blotting. Effects of OTUD3 on the cell growth were examined by colony formation assay (scale bar, 1 cm). Results are representative of n = 3 independent experiments. b OTUD3 WT and C76A were overexpressed in H1299 cells. The protein levels of OTUD3 WT and OTUD3 C76A were analyzed by western blotting. Effect of OTUD3 WT or C76A on the H1299 cells growth was examined by colony formation (scale bar, 1 cm). Results are representative of n = 3 independent experiments. c OTUD3 WT or OTUD3 C76A were overexpressed in human normal bronchial epithelium cells HBE. The protein levels of OTUD3 WT and OTUD3 C76A were analyzed by western blotting. Effects of OTUD3 WT or C76A on the cell growth were examined by colony formation assay (scale bar, 1 cm). Results are representative of n = 3 independent experiments. d OTUD3 was knocked down in lung cancer cell lines H1299 using shRNA. Effects of OTUD3 on the migration were examined by transwell assay (scale bar, 100 μm). Migrated cells were plotted as the average number of cells per field of view. Results are representative of n = 3 independent experiments. e Effect of OTUD3 WT or C76A on the migration were examined by transwell assay (scale bar, 100 μm). Migrated cells were plotted as the average number of cells per field of view. Results are representative of n = 3 independent experiments. f H1299 cells with or without OTUD3 knockdown were subcutaneously injected (2 × 10⁶ cells per mouse) into 6-week-old nude mice (n = 5). Tumor volume and weights were analysed (scale bar, 0.5 cm). g H1299 cells with or without overexpressing OTUD3 were subcutaneously injected (1 × 10⁶ cells per mouse) into 6-week-old nude mice (n = 5). Tumor volume and weights were analysed (scale bar, 0.5 cm). h A549 cells with or without OTUD3 knockdown were subcutaneously injected (2 × 10⁶ cells per mouse) into 6-week-old nude mice (n = 5). Tumor volume and weights were analysed (scale bar, 0.5 cm). i, A549 cells with or without overexpressing OTUD3 were subcutaneously injected (1 × 10⁶ cells per mouse) into 6-week-old nude mice (n = 5). Tumor volume and weights were analysed (scale bar, 0.5 cm). For a–e, the data are shown as mean ± s.d. Student’s t-test. For panels f–i, tumor volume was analyzed by two-way ANOVA test, tumor weight was analyzed by Student’s t-test. Statistics source data can be found in Supplementary Data 1. Uncropped images of blots are shown in Supplementary Fig. 9