GC-MS Analysis of siAd4BP/SF-1 Knockdown

Y-1 cells were treated with siAd4BP/SF-1 (#1635) or siControl for 48 h, and then whole cells or mitochondria isolated from the cells were suspended in 0.2 ml methanol at 4 °C. The suspension was sonicated (2 × 20 s pulses, separated by a 30-s interval) on a Bioruptor^® Plus (Diagenode). Thereafter, they were centrifuged at 15,000 rpm for 5 min at 4 °C to eliminate methanol-insoluble cellular components. The supernatant (methanol soluble fraction) was recovered and evaporated to remove methanol. Gas chromatography-mass spectrometry analysis was performed using an Agilent 6890 Plus gas chromatograph interfaced with a single-quadrupole Agilent 5975C MSD (Agilent) as previously described^{49 (link)}.