For Western blotting, skeletal muscle tissues were homogenized with RIPA buffer. After the lysates were centrifuged at 13,000 rpm for 30 min, the supernatants were measured using bicinchoninic acid (BCA) (Thermo Fisher Scientific, Waltham, MA, USA) assay and boiled for 5 min at 100 °C after mixing with a 5× sample buffer. Western blotting samples were loaded onto SDS-PAGE gels, run, and transferred onto PVDF membranes (Merck Millipore, Burlington, MA, USA). After blocking with Tris Buffered Saline with Tween 20 (TBST) containing 5% skim milk, the membranes were incubated with primary antibodies overnight at 4 °C and then incubated with secondary antibodies for 1–2 h at RT. The membranes were incubated with ECL solution (Promega, Madison, WI, USA) and imaged using ImageQuant LAS 500 (GE Healthcare, Chicago, IL, USA).
The primary antibodies used were anti-BIS [1 (link)], anti-HSF1, anti-HSP70 (Enzo Life Sciences), anti-GAPDH, anti-HSPB5, anti-YAP1 (Santa Cruz Biotechnology), anti-cleaved PARP1, anti-desmin, anti-HSPB8, anti-p62 (ABCAm), anti-COX4 (Cell Signaling Technology, Danvers, MA, USA), and anti-filamin C (Novus Biologicals, Centennial, CO, USA).
The primary antibodies used were anti-BIS [1 (link)], anti-HSF1, anti-HSP70 (Enzo Life Sciences), anti-GAPDH, anti-HSPB5, anti-YAP1 (Santa Cruz Biotechnology), anti-cleaved PARP1, anti-desmin, anti-HSPB8, anti-p62 (ABCAm), anti-COX4 (Cell Signaling Technology, Danvers, MA, USA), and anti-filamin C (Novus Biologicals, Centennial, CO, USA).