Fig. 6: EGFR protein stabilization and deposition at the endoplasmic reticulum membrane is mediated by LAMC2.

A Western blotting analyses indicating membrane EGFR protein expression levels in LAMC2 knockdown or overexpressing lung cancer cell lines. B EGFR membrane luminescence measured by flow cytometry. *p < 0.05, **p < 0.01. C EGFR protein levels found in the membrane and whole cell lysates (WCL) of control and LAMC2 knockdown A549 and HCC827 cells, as well as LAMC2 overexpressing A549 cells subjected to BFA treatment. Quantification of inhibition of EGFR membrane localization, right. D Western blotting analyses indicating protein levels of EGFR and LAMC2 in control and LAMC2 knockdown A549 and HCC827 cells, as well as LAMC2 overexpressing A549 cells subjected to BFA treatment. Quantification of EGFR half‐life changes, right. E Representative images of immunofluorescence staining of DAPI, LAMC2, and GRP78 showed co-localization between LAMC2 and GRP78 at the nuclear/ER membrane. F Representative images of immunofluorescence staining of DAPI, EGFR, and GRP78 in NC and LAMC2 overexpressing A549 cells subjected to BFA treatment. G A549 cells were transfected with LAMC2-His and subjected to CHX treatment (10 μg/ml). Protein extracts were subjected to co-immunoprecipitation, ubiquitination, and immunoblot assays. H A549 cells were transfected with LAMC2-His and subjected to BFA treatment. Protein extracts were subjected to co-immunoprecipitation, ubiquitination, and immunoblot assays. I A549 LAMC2 knockdown cells were subjected to BFA treatment. Protein extracts were subjected to co-immunoprecipitation, ubiquitination, and immunoblot assays. J A549 cells were subjected to LAMC2 knockdown, BFA, and/or MG132 (1 μM) treatments. Western blotting analyses were performed to assess protein levels of EGFR and LAMC2.