Fig. 4: The Dsg2 IL domain directly bound the cSrc SH2 domain. | Cell Death & Differentiation

Fig. 4: The Dsg2 IL domain directly bound the cSrc SH2 domain.

From: Loss of desmoglein-2 promotes gallbladder carcinoma progression and resistance to EGFR-targeted therapy through Src kinase activation

Fig. 4

a Immunoprecipitation (IP) and immunoblot (IB) analyses for Dsg2, cSrc, FAK, Akt, and ERK1/2 in GBC cells. WCL whole-cell lysates; IgG1 normal rabbit immunoglobulin G1. b The JCRB1033 cells were transfected with 10 μM cSrc or FAK siRNA for 48 h. Cell lysates were immunoprecipitated with an anti-Dsg2 antibody, blotted, and probed with an anti-cSrc or anti-FAK antibody. c GBC cell lysates IP with an anti-cSrc antibody and probing with indicated antibodies. d Co-IP analysis was performed to determine interactions between Dsg2 and Src pTyr527 in GBC cells. e Schematic diagram of the eGFP-tagged cSrc protein and the mutant constructs. f Western blot analysis of JCRB1033 lysates following IP with an anti-Dsg2 antibody after expressing the indicated constructs. Cell lysates were immunoprecipitated with an anti-Dsg2 antibody, blotted, and probed with an anti-cSrc antibody. g Diagram showing the strategy used to construct plasmids used to express FLAG-tagged hDsg2 truncation mutants. h Western blot analysis of the expressed hDsg2 variants using an anti-FLAG tag antibody (top left blot) and after lysate IP with an anti-cSrc antibody and probing with an anti-FLAG-tagged antibody (top right blot). i The eGFP-cSrc construct and each hDsg2 fragment were co-transfected into JCRB1033 cells. Western blots following anti-GFP IP of the indicated Dsg2 variants from cell lysates. Cyto cytoplasmic domain, IA intercellular anchor domain, ICS intercellular cadherin-like sequence domain, IL intercellular linker domain.

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