*4.5. Western Blotting*

Protein was obtained by lysing tissue or cells in radioimmunoprecipitation assay (RIPA) buffer (NEB, #9806). Protein concentration was measured using PierceTM BCA Protein Assay Kit (Thermo Scientific, Waltham, MA, USA; #23225) on an iMarkTM Microplate Reader (BioRad, Hercules, CA, USA). Proteins were electrophoresed in a polyacrylamide (10%) gel (Roth, Karlsruhe, Baden-Wuerttemberg, Germany; #3029.1) and transferred onto a Roti® polyvinylidene fluoride (PVDF) membrane (Roth, Karlsruhe, Baden-Wuerttemberg, Germany; #T830.1). Membranes were blocked, stained with primary antibodies, and horseradish peroxidase (HRP)-coupled secondary antibodies. Antibodies and concentrations are given in Supplementary Materials Table S2. PierceTM ECL Western Blotting Substrate (Thermo Scientific, Waltham, MA, USA; #32106) and ChemiDocTMMP (Biorad, Hercules, CA, USA) were used for detection of stained proteins.
