P616 drp1

Proteins were extracted from isolated islets or NIT-1 cells using RIPA lysis buffer. Proteins (15–20 μg) were separated by SDS-PAGE and transferred onto nitrocellulose membranes (Whatman, Clifton, NJ, USA). After incubation with specific antibodies, the membrane bands were visualized by Amersham Imager 680 Blot and Gel Imagers (GE Healthcare Life Sciences, Marlborough, MA, USA). Antibodies against SENP2 (Santa Cruz Biotechnology Inc.), p616 DRP1 and DRP1 (Cell Signaling Technology, Danvers, MA, USA), GFP/YFP (Arigo Biolaboratories, Hsinchu City, Taiwan), GAPDH (Merck Millipore, Burlington, MA, USA), tubulin and Flag (Sigma-Aldrich) and OXPHOS Rodent WB antibody cocktail (Abcam, Cambridge, UK) were used for western blotting.

Antibodies directed against the following were used in this study: SLC25A46 (Proteintech, 12277‐1‐AP), MFN2 (Santa Cruz, sc‐100560), SDHA (Abcam, ab14715), AIFM1 (Chemicon, MABN92), ATP5A1 (Abcam, ab14748), HCCS (Proteintech, 15118‐1‐AP), SCO1 (in‐house), MRPL14 (Proteintech, 15040‐1‐AP), NDUFA9 (Abcam, ab14713), UQCRC2 (Abcam, ab14745), COX4 (in‐house), COX2 (in‐house), MFN1 (Cell Signaling, 14739), IMMT/MIC60 (Abnova, H00010989‐M01), CHCHD3/MIC19 (Aviva, ARP57040‐P050), OPA1 (BD biosciences, 612607), MFF (Sigma, SAB1305258), CDKN1A/p21 (Cell Signaling, 2946), p53 (Abcam, ab176243), p‐Mdm2 (Cell Signaling, 3521), LAMP1 (Santa Cruz, sc‐18821), VIM (BD biosciences, 550513), VDAC1 (Abcam, ab14734), EMC1 (GeneTex, GTX119884), EMC2 (ProteinTech, 25443‐1‐AP) and FlagM2 (Sigma, F1804), TOMM20 (Santa Cruz, sc‐11415), DRP1 (BD biosciences, 611113), p637‐DRP1 (Cell Signaling, 6319), p616‐DRP1 (Cell Signaling, 3455), KDEL (Abcam, ab50601), ITPR1 (Cell Signaling, 8568), alpha‐tubulin (Santa Cruz, sc‐23948).