N methyl 14c sm

[N-methyl-¹⁴C] SM (52 mCi/mmol) was purchased from PerkinElmer (USA). Biotin-SM was purchased from Echelon Biosciences Inc. (USA). Streptavidin-Sepharose High-Performance Beads were purchased from GE Healthcare Bio-Sciences AB (Sweden). Percoll and Mono Q anion exchange columns and Superose 6 and 12 gel filtration columns were purchased from Amersham Biosciences (UK). Anti-nSMase2, anti-SMase1, and anti-SNAP25 mouse monoclonal antibodies and anti-c-Myc antibodies were purchased from Santa Cruz Biotechnology (USA). Anti-Hsp60 mouse monoclonal antibody was purchased from Stressgen Biotechnologies (USA). Anti-VDAC and anti-COX IV rabbit monoclonal antibodies were purchased from Cell Signaling Technology (USA). Ethanol (absolute GR for analysis) was obtained from Merck (Germany). Purified Mouse Anti-cytochrome c antibody was purchased from BD Biosciences (USA). A Cytosol/Mitochondrial Fractionation kit was purchased from Calbiochem (Germany). Mini-PROTEAN^® TGX^TM Gels (4%-15%, 15-well comb, 15 μl) were purchased from Bio-Rad Laboratories (USA). Unless otherwise stated, all reagents used in this study were of the highest purity and were purchased from Sigma-Aldrich (USA). Male Sprague–Dawley rats (5 to 7 weeks old) were purchased from Orient Bio (Korea). All animal experiments were approved by the Chung-Ang University Institutional Animal Care and Use Committee (IACUC No. 201700061).