Anti col3a1

Tensile tissue or treated fibroblasts were lysed in radioimmunoprecipitation buffer (RIPA) containing protease and phosphatase inhibitors as previously described (Jiang et al., 2014 (link)). A BCA Protein Assay Kit (Thermo Fisher Scientific, IL, USA) was employed to determine protein concentration. Twenty micrograms of lysate were loaded onto a 10 to 15% sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) for separation and then electrotransferred to a polyvinylidene fluoride membrane (PVDF; Millipore, Bedford, Massachusetts, USA). Membranes were incubated with primary including anti-COL3A1, anti-α-SMA, anti-ATF-6, anti-phospho-IRE1 (anti-p-IRE1), anti-CHOP, anti-Bcl-2 (Abcam, Cambridge, MA, USA), anti-GRP78 (glucose regulated protein 78), anti-Bax, and anti-β-actin (Cell Signaling Technology, Danvers, MA, USA) and secondary antibody (Abcam, Cambridge, MA, USA). β-Actin was used for standardization. A semi-quantitative analysis was performed using ImageJ software.