Figure 3.
Effects of XCT-790 on mitochondrial functions in 3T3-L1 adipocytes. Expression levels of PDK2 and PDK4 were determined by RT-PCR. Relative expression levels were normalized with GADPH and presented as the fold of DMSO (A). SDH activity was determined by MTT method. Absorbance was read at 570 nm The relative activity of SDH was presented as the fold of control, values were shown as mean ± S.D. of triplicate (B); Mitochondrial membrane potential was analyzed by calculating the ratio of cells with red and green fluorescence generated by JC-1. Cells with more red fluorescence mean higher membrane potential. Fluorescence of harvested adipocytes was detected by FACs. CCCP, which uncouples the membrane potential serves as the positive control. PE: red fluorescence FITC: green fluorescence (C); Total cellular ATP levels were measured by ATPlite-glo, a luciferase-based luminescence assay kit (PerkinElmer) (D); ROS levels were determined by measuring the fluorescence of DCF with FACs (excitation at 488 nm, emission at 530 nm). The relative levels are presented as the fold of DMSO with Rose up as a positive control. Values were shown as mean ± S.D. of triplicate (E).