Anti glyceraldehyde 3 phosphate dehydrogenase gapdh 14c10

Q dehydrate, H₂O₂, and anti-Cav-1 (c3237) were purchased from Sigma Aldrich Co. (St. Louis, MO). Anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 14C10) and anti-phospho-Fyn (D49G4) were obtained from Cell Signaling Technology (Beverly, MA). Anti-phospho-Cav-1 (sc-14037) and anti-Fyn (sc-16) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Q3GA and myricetin (M) (1127S) were obtained from Extrasynthese (Genay, France). Flavone (F) was obtained from Nacalai Tesque (16012-31) (Kyoto, Japan). Luteolin (L) was obtained from LKT Laboratories (L8377; Saint Paul, MN) and 8-prenyl Q was synthesized as previously described by Kawamura et al.^{(40 )}

After collection in ice‐cold PBS, cells were lysed in RIPA buffer containing 1 mM DTT, and protease and phosphatase inhibitors (89900; Thermo Fisher Scientific). The soluble fraction was subjected to SDS‐PAGE (456‐1086; Bio‐Rad, Hercules, CA) and protein transfer was performed using a Trans‐Blot Turbo Transfer System (1704156; Bio‐Rad) or Blocking One (03953‐95; Nakarai, Kyoto, Japan). The following primary antibodies were used: anti‐dysferlin (1:1,000, ab124684), anti‐glyceraldehyde 3‐phosphate dehydrogenase (GAPDH; 14C10; 1:1,000–3,000, 2118; Cell Signaling Technology, Danvers, MA), anti‐p62 (1:1,000, 66184‐1‐JG; Proteintech, Rosemont, IL), and anti‐LC3 (1:1,000, M186‐3; MBL, Nagoya, Japan). Secondary antibodies included donkey Horse Radish Peroxidase (HRP)‐linked anti‐rabbit IgG (NA934‐1ML) and sheep HRP‐linked anti‐mouse IgG (NA931‐1ML), both from GE Healthcare, Little Chalfont, U.K. Antibodies were diluted in Canget Signal solution (NKB‐101; TOYOBO, Osaka, Japan) and blotting was performed using the iBind Western System (Thermo Fisher Scientific) at room temperature. Immunoblot detection was performed using ChemiDoc Touch Imaging System (Bio‐Rad) and band area was quantified by Image Lab (Bio‐Rad).