Fig. 4

p110α Y317F mutation attenuates Src-MLC2 signaling pathway. A The bubble diagram of GO analysis with differentially phosphorylated proteins between HCT116 parental cells and HCT116 Y317F KI mutant cells. B HCT116 parental cells and HCT116 Y317F KI mutant cells were serum-starved overnight and then treated with EGF or insulin for 15 min. Cell lysates were collected and blotted with indicated antibodies. C-F MLC2 reconstitution rescues the proliferation and migration defects caused by p110α Y317F mutation. MLC2 was overexpressed in HCT116 p110α Y317F KI mutant cells. p-MLC2 and total MLC2 protein levels were evaluated by Western blots (C). The asterisk indicates exogenously overexpressed MLC2. Cell lines including HCT116, HCT116 Y317F KI mutant, HCT116 Y317F KI mutant reconstituted with MLC2 were analyzed with for proliferation (D); colony formation (E); and migration (F). Two-tailed unpaired t test (E, F) and two-way ANOVA (D), *p < 0.05, **p < 0.01, ****p < 0.0001, ns, not significant