Fig. 4
From: SUMOylation of AnxA6 facilitates EGFR-PKCα complex formation to suppress epithelial cancer growth
AnxA6 SUMOylation inhibits activation of EGFR mutation-induced signal transduction. A The the silencing of AnxA6 activate pY-GFP-EGFRT790M/L858R and pERK1/2. HeLa stable cells were generated by using lentiviral infections with indicated plasmids, and cells were transfected with pGFP-EGFRT790M/L858R plasmids for 24 h, starved overnight and followed with/without 10 ng/ml EGF treatment for 3 min. Cell lysates were immunoblotted with the indicated antibodies. B The K299R mutation of AnxA6 impaired its ability to inactivate pY-EGFRT790M/L858R and pERK1/2. A431 cells were co-transfected with the indicated plasmids for 24 h, starved overnight and followed with/without 10 ng/ml EGF treatment for 3 min. Cell lysates were immunoblotted with the indicated antibodies. C The K299R mutation impaired its recruitment ability of AnxA6 to promote GFP-EGFRT790M/L858R-PKCα interaction. A431 cells were co-transfected with the indicated plasmids for 24 h, starved overnight and followed with/without 10 ng/ml EGF treatment for 3 min. Cell lysates were IP to capture GFP-tagging EGFRT790M/L858R and subsequently immunoblotted with indicated antibodies. D The K299R mutation of AnxA6 impaired its ability to interact GFP-EGFRT790M/L858R. A431 cells were co-transfected with the indicated plasmids for 24 h, starved overnight and followed with/without 10 ng/ml EGF treatment for 3 min. Cell lysates were IP to capture Flag-tagging AnxA6 and subsequently immunoblotted with indicated antibodies. IP: immunoprecipitation, IB: immunoblot, Input: same account of cell lysate to load