Fig. 5
PHB-knockdown impairs STOML2-induced CRC proliferation and tumor growth. a Immunoblot analysis of STOML2, PHB and MAPK signaling pathway in SW480-STOML2 cells, with/without PHB knockdown. b Cell-cycle distribution analysis of STOML2-overexpressed PHB-knockdown SW480 and control. Fraction of cells in G1/G2/S phase was shown. c CCK8 proliferation assay. Indicated cells were seeded 5000/well in 96-well culture-plates and OD values were measured by day. Data are presented as means ± SEM from five independent experiments. *P < 0.05, **P < 0.01, two-way ANOVA. d Colony formation of indicated cells. 5000 indicated cells were seeded 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-k Subcutaneous implantation of MC38-STOML2-scr and MC38-STOML2-shPHB cells in C57BL/6 mice. (f) Tumor volume (cm3) measured at indicated time points between two groups. *P < 0.05, two-way ANOVA. (g) 2 × 106 cells were injected into hind limbs of each group; tumors were retrieved at 21st day after injection. (h) Tumor weight (g) measured at retrieval. **P < 0.01, t test. (i) Representative image of HE staining of subcutaneous implant in each group. Scale bar, 40 μm. (j) Representative images of IHC staining show negative correlation between Ki-67 and PHB knockdown in each group. Scale bar, 40 μm. (k) Relative intensity of Ki-67 staining, ***P < 0.001, t test