Fig. 2.

PPARγ upregulates ATGL independently of SIRT1 expression. A: Control (EV) and SIRT1-ablated (Sh) 3T3-L1 adipocytes were transfected with 30 nM of scrambled (sc) RNA or siRNA directed against mouse PPARγ (si-Pγ) on day 8 of differentiation. After 48 h, cells were homogenized and total lysates (50 µg) were analyzed by Western blotting. B: Relative band intensities for ATGL (gray bars), HSL (white bars), and PPARγ (black bars) were determined in three independent experiments and normalized by cellugyrin. C: Control (EV) and SIRT1-ablated (Sh) 3T3-L1 adipocytes were transfected with scrambled (sc, black bars) RNA or siRNA directed against mouse PPARγ (si-Pγ, gray bars). Glycerol was measured in media aliquots without (basal) or with (stimulated) 10 μM isoproterenol (Iso) for 2 h. A representative result of three independent experiments is shown. Data are presented as nM glycerol/mg of protein/h and expressed as mean ± SD. NS, not significant. * P < 0.05; ** P < 0.001.