FIGURE 7.

Adipocyte functions in Tfam knockdown cells. A, the relative mRNA level of various transcription factors and adipocyte-specific proteins was determined by qRT-PCR at day 5 in cells transfected at day 2 with scrambled or Tfam-directed siRNA oligonucleotides. Results are expressed relative to scrambled values. B and C, Oil red O staining and triglyceride content of cells at day 5. D, cells were washed in PBS and placed in fresh medium. An aliquot of the medium was collected at the time points indicated and analyzed by Western blotting with an antibody to adiponectin (Acrp30). E, cells were incubated without or with 50 ng/ml TNFα for 3 h, and reactive oxygen species were measured by 5-(and 6-)chloromethyl-2',7'-dichlorodihydroflioresceindiacetate, acetyl ester fluorescence, detected directly in cells grown on 24-well multiwell plates using a TECAN Safire² microplate reader.