Mitochondrion Isolation
Mitochondria isolation was performed using the commercial kit (Mitochondria Isolation Kit for Cultured Cells, Thermo Fisher Scientific, Cat. No. 89874) according to the manufacturer’s protocol. In brief, 50 million cells pre-treated with 4-OH tamoxifen for 24 h were harvested and collected in reagent A, followed by 2-min incubation on ice. Cells were then treated with 10 μL of reagent B and incubated for 5 min on ice with vertexing every minute. After adding the reagent C, homogenized cells were centrifuged at 700 g for 10 min at 4 °C and the pellet was discarded. The crude fraction was centrifuged at 12,000 ×