Anti total oxphos cocktail

Cells or tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer (30 mM Tris [pH 7.5], 150 mM sodium chloride, 1 mM sodium phenylmethylsulphonyl fluoride, 1 mM sodium orthovanadate, 1% Nonidet P-40, 10% glycerol, and phosphatase and protease inhibitors) containing a 1x protease inhibitor cocktail. Western blotting was performed with protein (30–50 μg) using commercially available antibodies: anti-SHMT2 (#12762, 1:1000, Cell signaling Technology, Beverley, MA, USA), anti-MTHFD2 (ab151447, 1:1000, Abcam, Cambridge, USA), anti-total OXPHOS cocktail (ab110413, 1:1000, Abcam), anti-β-actin (ab8227, 1:1000, abcam), and anti-α-tubulin (T5168, 1:1000, Sigma-Aldrich). The appropriate secondary antibodies were obtained from Bio-Rad (Hercules, California, USA). Images were acquired using the Odyssey imaging system and quantified using Image Studio Digits (LI-COR Biosciences, Lincoln, NE, USA).

Western blotting analysis was performed as previously described [20 (link)]. Antibodies used in this study are as follows: anti-SMYD1 (Santa Cruz, SC2059 and Abcam, ab32482) Anti-FLAG (Sigma Aldrich F1804), Anti-SMYD2 (Abcam, ab108217), Anti-SMYD4 (Thermo Fisher Scientific, PA5-96,631), Anti-HSP90 (Cell Signaling Technology, 4874S), Anti-p53 (Abcam, ab131442), Anti-PGC-1α (Abcam, ab54481), Anti-Histone H3 (Abcam, ab1791), Anti-H3 tri methyl K4 (Abcam, ab8580), Anti-beta TUBULIN (ab6046), Anti-ACTIN (sc-1616), Anti-NDUFS3 (Abcam, ab196019), Anti-NDUFS2 (Abcam, ab110249), Anti-NDUFV1 (GeneTex, GTX102209), Anti-NDUFV2 (Proteintech, 15,301-1-AP), Anti-NDUFA9 (Abcam, ab14713), Anti-NDUFA10 (Santa Cruz, SC-376357), Anti-total OXPHOS cocktail (Abcam, ab110413) and Anti-VDAC1 (Abcam, ab14734), Anti-OPA1 (BD Biosciences, 612,606), Anti-MFN2 (Abcam, ab50843), Anti-Mitofilin (Proteintech, 10,179-1-AP).

Anti-PKA Catα (C-20) (sc-903; Santa Cruz Biotechnologies, Dallas, TX, USA), anti-PKA Reg I α/β (G-6) (sc-271446; Santa Cruz Biotechnologies), anti-PKA Reg II (L-16) (sc-26803; Santa Cruz Biotechnologies), anti-phospho-PKA substrate (RRXS*/T*)(100G7E) (#9624; Cell Signaling Technology, Danvers, MA, USA), anti-PP1 (E-9) (sc-7482; Santa Cruz Biotechnologies), anti PP2A α/β (ab32141; Abcam, Cambridge, UK), anti-LC3 I/II antibody (ABC929; Merck KGaA), anti-calreticulin (#2891S; Cell Signaling Technology), anti-GAPDH (sc-166574; Santa Cruz Biotechnologies), anti-total OXPHOS cocktail (ab110413; Abcam), anti-ATP synthase subunit α (ab14748; Abcam), and anti β-actin (A3854; Merck KGaA). Peroxidase-conjugated secondary antibodies were purchased from Merck or Santa Cruz Biotechnologies.