Fig. 2
STOML2 promotes CRC cell proliferation and colony formation in vitro. a Cell-cycle distribution analysis of STOML2-knockdown SW620 and control. Fraction of cells in G1/G2/S phase was shown. b Immunoblot of STOML2 and GAPDH in SW620 transfected with STOML2-knockdown lentivirus (SW620-shSTOML2) and control group (SW620-scr). c CCK8 proliferation assay. Indicated cells were seeded 5000 per well in 96-well culture-plates and OD values were measured by day. Data are presented as means ± SEM from five independent experiments. ***P < 0.001, two-way ANOVA. d Colony formation of SW620-shSTOML2 and -scr. Indicated cells were seeded 5000 per well in 6 wells culture-plates. Giemsa staining was carried out after 14 days of culture. e Number of colonies formed on each plate. Data presented as means ± SEM from three independent experiments. ***P < 0.001, **P < 0.01, one-way ANOVA. f Immunoblot assay of indicated protein in SW480 transfected with STOML2-overexpressed lentivirus (SW480-STOML2) and control group (SW620-mock). g Cell-cycle distribution analysis of STOML2-overexpressed SW480 and control. Fraction of cells in G1/G2/S phase was shown. h CCK8 proliferation assay. Indicated cells were seeded 5000/well in 96-well culture-plates and OD values were measured by day. Data was presented as means ± SEM from five independent experiments. ***P < 0.001, two-way ANOVA. i Colony formation of SW480-STOML2 and -mock. 5000 indicated cells were seeded in 6 wells culture-plates. j Number of colonies formed on each plate. Data presented as means ± SEM from five independent experiments. *P < 0.05, t test. k-n Organoid culture, CCK8 assay and IF staining of STOML2-overexpressed organoid derived from intestinal tumor of APCMin/+ mice (APCmin-STOML2) and control group (APCmin-mock). (k) Representative images of indicated groups of organoids on first and tenth day of culture. Scale bar, 200 μm. (l) Calculated surface area (μm2 × 103) of indicated groups of organoids. (m)CCK8 proliferation assay of organoids. Data presented as means ± SEM from three independent experiments. ***P < 0.001, two-way ANOVA. (n) Separated and merged images of immunofluorescent staining to detect Ki-67 (red), E-cadherin (green), DAPI (blue) in indicated organoids (scale bar, 20 μm)