Uqcrc1

Antibodies list: CPT1A (A5307, ABclonal, Wuhan, China), UCP1 (A5857, ABclonal), UCP3(A16996, ABclonal), PPARα (sc-9000, Santa Cruz, Dallas, TX, USA), PPARγ (2435S, CST, Danvers, MA, USA), FAS (3189S, CST), SREBP1 (Sc13551, Santa Cruz), PGC-1α (TA319007, Origene, Rockville, MD, USA), NDUFS3 (459130, Invitrogen, Waltham, MA, USA), SDHB (459230, Invitrogen), UQCRC1 (459140, Invitrogen), COX4 (459600, Invitrogen), ATP Synthase Subunit Alpha (459240, Invitrogen), SOD2 (sc-137254, Santa Cruz), β-Actin (3700S, CST), α-Tubulin (3873S, CST).
Mice diet was provided by SLAC Laboratory Animal Co. Ltd. (Shanghai, China). Insulin was purchased from Nove Nordisk A/S. The blood glucose meter and blood glucose test strip were both purchased from ROCHE (ACCU-CHEK Active).
The Reverse Transcription System kit was purchased from Promega; SYBR green was purchased from Takara; polymerase chain reaction (PCR) primers were synthesized by Beijing Qingke biotechnology Co. Ltd. Nitrocellulose membranes used in W.B. were purchased from PerkinElmer Life Sciences. Other reagents used in this study were purchased from Sigma (St. Louis, MO, USA).

About 10⁶ culture fibroblasts from patient and controls were pelleted and incubated on ice with digitonin 0.4% for 10 min. Then samples were washed twice with PBS and incubated on ice with dodecylmaltoside 1% and NativePAGE Sample Buffer 1X (Invitrogen) for 15 min. After the incubation, samples were centrifuged at 20,000 × g for 30 min and the supernatants were measured out. Then 10 μg of supernatants plus the NativePAGE 0,25% G-250 sample additive (Invitrogen) were separated by 3–12% gradient NativePAGE Novex Bis-Tris Gels (Invitrogen) and transferred to nitrocellulose membranes and incubated with antibody against UQCRC1 (Invitrogen), NDUFS9 (Invitrogen) and SDHA (Invitrogen).