Mitosox detection kit

Antibodies against Drp1, p62, LC3, RIP1, RIP3, TSG101, and CD63 were purchased from Abcam (Cambridge, MA, USA). β-actin, tubulin, and COX IV, which were used as references for the total, cytoplasmic, and mitochondrial fractions, were also acquired from Abcam (Cambridge, MA, USA). Antibodies against Phospho-RIP3 (Thr231/Ser232) and Phospho-Drp1 (Ser616) were purchased from Cell Signaling Technology (Danvers, MA, USA). The Aggresome Detection Kit was acquired from Abcam (Cambridge, MA, USA). MitoTracker Deep Red, the ROS 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) kit, dihydroethidium (DHE) assay kit, and MitoSOX detection kit were purchased from Invitrogen (Carlsbad, CA, USA). Ad-mRFP-GFP-LC3 was supplied by Beyotime (Shanghai, China). Adenoviral vectors for Drp1 short hairpin RNA (shRNA) and Drp1 mutations (Drp1-S616D and Drp1-S616A) were generated by Genechem Technology (Shanghai, China). Related activators and inhibitors, including Mitochondrial division inhibitor 1 (Mdivi-1), N-acetylcysteine (NAC), and Necrostatin-1 (Nec-1), were obtained from Selleck (Shanghai, China). The Protein A/G Magnetic Beads IP Kit was purchased from Thermo Scientific (Waltham, MA, USA), and fetal bovine serum (FBS) and penicillin/streptomycin were procured by Invitrogen (Carlsbad, CA, USA). All other chemicals were supplied by Sigma unless otherwise specified.

Deoxyarbutin (4-[(tetrahydro-2H-pyran-2-yl) oxy] phenol, purity >99%) was purchased from Sigma (St. Louis, MO, USA). Culture medium (RPIM 1640 medium, DMEM medium), fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (100 mg/mL) were purchased from GIBCO (Life Technologies, Carlsbad, CA, USA). Hoechst 33258, Annexin V/PI apoptosis kit, Cell arrest detection kit, ATP detection kit and BCA Protein assay kit were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Mito-sox detection kit, JC-1 detection kit, Mito-tracker and ER-tracker were purchased from Invitrogen Life Technology (Grand Island, NY, USA). 5-FU and molecular probes dansyl chloride (DNS) were purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Unless indicated otherwise, the other reagents were purchased from Sigma. dA was dissolved in ethanol and diluted with fresh medium to achieve the desired concentration. The final concentration of ethanol did not exceed 0.1% in the fresh medium.