Fig. 5

Suppression of CRC progression in vitro and in vivo by MAGI3 is dependent on c-Myc. a, To overexpression c-Myc in CRC-MAGI3 cells by exogenous transfection of c-Myc. c-Myc was overexpressed in HT115-MAGI3 or RKO-MAGI3 cells by transfected with c-Myc constructs. b, MAGI3 had less effect on the CRC cells proliferation when c-Myc protein levels were overexpressed. Cell viability of HT115 and RKO cells by CCK-8 assay. ***P < 0.001 vs. pcDNA3.0 + Vector; ^###P < 0.001 vs. MAGI3 + Vector. c, Overexpressed c-Myc effectively restricted MAGI3-inhibition of colony formation of CRC cells. d, MAGI3 overexpression inhibited the growth of RKO xenografts in nude mice. Representative images of tumors from the implanted mice (left). The volume and weight of tumors were significantly decreased in MAGI3 overexpressed RKO xenograft tumors (middle and right). e, MAGI3 overexpression correlated with the reduced the expression of c-Myc in RKO xenografts. Representative IHC staining of MAGI3 and c-Myc in xenografted tumor. Scale bars: 50 μm. Dot plot (right) showed the corresponding quantification of MAGI3 or c-Myc H-score. f, Protein levels of MAGI3 were reduced coordinately with c-Myc increase in AOM/DSS induced CRC. Images of AOM/DSS induced CRC (top). The protein levels of MAGI3 and c-Myc in colorectal tumor (T) and matched surrounding tissues (S) were detected by western blot (bottom). Data were presented as the mean ± SEM. Statistical significance was determined by 2-way ANOVA with Bonferroni post-tests (b and d-middle), 2-tailed, unpaired t test (c and d-right) and Mann–Whitney test (e). *P < 0.05, **P < 0.01, ***P < 0.001