Normal rabbit igg for immunoprecipitation

MG132 was obtained from Enzo Life Science (NY), and 17β-estradiol (E₂) was purchased from Sigma-Aldrich (St. Louis, MO). An antibody against α-ENaC was obtained from GenScript, as previously described^{19 (link),20 (link)}. Antibodies against derlin-1 and HA-tag were purchased from Sigma-Aldrich. Antibodies specific for ubiquitin, p-AMPK, and AMPK were purchased from Cell Signaling Technology (Danvers, UK). β-Actin, α-tubulin, and GAPDH antibodies were purchased from Proteintech (Chicago, IL). Other antibodies, including normal rabbit IgG for immunoprecipitation and secondary mouse or rabbit antibodies for western blotting, were purchased from Thermo Fisher (Waltham, UK). shRNA-derlin-1 and shRNA-scramble constructs were designed and cloned into the pcDNA3.1 vector. HA-tagged WT-AMPK-α1 or DN-AMPK, a dominant-negative (DN) AMPK-α1-K45R mutant, were gifts from Dr. Kenneth Hallows (University of Southern California). Wild-type ubiquitin was amplified by PCR and cloned into pcDNA3.1. All plasmid constructs were confirmed by DNA sequencing.

Antibodies against S100A16, myosin-9, E-cadherin, N-cadherin, vimentin, collagen I, and GAPDH were purchased from Proteintech (Chicago, USA). Antibodies against fibronectin and α-SMA were purchased from Sigma-Aldrich (St Louis, MO). Other antibodies, including normal rabbit IgG for immunoprecipitation and secondary mouse or rabbit antibodies for western blotting, were purchased from Thermo Fisher (Waltham, UK). TGF-β was purchased from R&D (Minneapolis, USA). Rhod-2 AM and BAPTA-AM were purchased from Sigma-Aldrich (St Louis, MO). The constructs of shRNA-S100A16 and shRNA-scrambled were designed and cloned into pcDNA6.2. All plasmid constructs were confirmed by DNA sequencing.