ANT2 siRNA (m)

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	3T3-L1 preadipocytes were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), at 10% CO2 and 37°C. Differentiation of 3T3-L1 cells was induced as described previously 48. Briefly, confluent cells were incubated for 2 days in a medium comprising DMEM supplemented with 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine, 1 μM dexamethasone, and 10 μg of insulin/ml. Thereafter, medium was replaced every other day with DMEM containing 10% FBS and 10 μg/ml insulin.The negative control (sc-37007) and specific siRNAs against ANT2 (sc-72506), and HIF-1α (sc-35562) were purchased from Santa Cruz Biotechnology, Inc. Transfections were performed with DharmaFECT 1 transfection reagent (Dharmacon) and conducted according to the manufacturer’s instruction

Protocol tips

3T3-L1 preadipocytes were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), at 10% CO2 and 37°C. Differentiation of 3T3-L1 cells was induced as described previously 48. Briefly, confluent cells were incubated for 2 days in a medium comprising DMEM supplemented with 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine, 1 μM dexamethasone, and 10 μg of insulin/ml. Thereafter, medium was replaced every other day with DMEM containing 10% FBS and 10 μg/ml insulin.The negative control (sc-37007) and specific siRNAs against ANT2 (sc-72506), and HIF-1α (sc-35562) were purchased from Santa Cruz Biotechnology, Inc. Transfections were performed with DharmaFECT 1 transfection reagent (Dharmacon) and conducted according to the manufacturer’s instruction

Publication protocol

3T3-L1 preadipocytes were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), at 10% CO2 and 37°C. Differentiation of 3T3-L1 cells was induced as described previously 48. Briefly, confluent cells were incubated for 2 days in a medium comprising DMEM supplemented with 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine, 1 μM dexamethasone, and 10 μg of insulin/ml. Thereafter, medium was replaced every other day with DMEM containing 10% FBS and 10 μg/ml insulin.The negative control (sc-37007) and specific siRNAs against ANT2 (sc-72506), and HIF-1α (sc-35562) were purchased from Santa Cruz Biotechnology, Inc. Transfections were performed with DharmaFECT 1 transfection reagent (Dharmacon) and conducted according to the manufacturer’s instruction

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