FIGURE 2.

PEDF induces expressions of TRAIL and PPARγ in BMDMs. A, BMDMs were treated with PEDF at the indicated concentrations for 24 h, and the cells were then processed for RT-PCR analysis. GAPDH expression was examined for normalization purposes. B, cells were treated as described above, and proteins were detected by Western blot analysis with antibodies as indicated. Representative blots (left panels) and densitometric analysis with S.D. (error bars) (right panels) of three independent experiments are shown. *, p < 0.05 versus untreated cells. C, BMDMs were exposed to PEDF for the time indicated and then harvested for Western blotting with antibodies against TRAIL (∼33.4 kDa) and PPARγ (∼57.6 kDa). Equal protein loading was confirmed by reprobing the membranes with a β-actin antibody. Representative blots and densitometric analyses with S.D. from four separate experiments are shown. D, PEDF-treated BMDMs were assessed for cell surface TRAIL expression. BMDMs were treated with PEDF or its solvent for 24 h. Cell membrane and cytosolic fractions were isolated as described under “Experimental Procedures” and then subjected to Western blot analysis. A representative result from two independent experiments is shown. E, BMDMs were treated with PEDF or solvent control for 24 h. The cells were then incubated with PE-conjugated isotypic control or PE-conjugated TRAIL antibody and analyzed by flow cytometry.