Fig. 7

Overexpression of COX4I2 promotes angiogenesis and CAF activation in CRC. a The HUVEC migration assay. b, c HUVEC migration analysis (Transwell assay) (magnification: ×200, scale bars: 50 μm). d The tube formation assay. e, f HUVEC tube formation and graph showing tube formation indices in different groups (magnification: ×200, scale bars: 50 μm). g Non-contact co-culture of MSCs and CRC cells. Cells were used in a 1:1 ratio. h, i Western blot showing CAF-related markers after 5 days of co-culture (n = 3 replicates). j–l Immunofluorescence of MSCs co-cultured with CRC cells (control cells and CRC cells transfected with the NC, sh-COX4I2, and oe-COX4I2 constructs) treated with or without 5 µM PD-166866 (magnification: ×400, scale bars: 20 μm). m The CAF migration assay. n, o Recruitment of CAFs by CRC cells (Transwell) (magnification: ×200, scale bars: 50 μm). Cell numbers were assessed using Image J. Data are expressed as the mean ± SEM. Asterisks indicate a significant difference at *P < 0.05, **P < 0.01, ***P < 0.001. All experiments were repeated at least three times, independently. For Abbreviations, see Abbreviation list