Figure 5.
Participation of PPARγ in iNOS inhibition by HP24. (A) Peritoneal macrophages from T. cruzi-infected mice (9 dpi) were obtained. Cells were transfected with PPARγ-siRNA during 72 h. Transfected or non-transfected cells were treated with HP24 (100 μM) for 48 h. iNOS expression was detected by immunofluorescence with a rabbit polyclonal anti-iNOS antibody and with a secondary goat anti-rabbit Alexa 488-labeled antibody. Cell nuclei were stained with DAPI. Mean fluorescence intensity (MFI) represents iNOS expression. Representative microphotographs are shown. Scale bar: 10 μm. (B) Western blot analysis was carried out and iNOS expression was determined. Protein levels were normalized against α-actin. (C) NO release to culture supernatants was analyzed by the Griess method. Results are expressed as the mean of three independent experiments (six mice/group). *P < 0.05 vs. uninfected cells, #P < 0.05 vs. T. cruzi-infected cells. ΔP < 0.05 vs. T. cruzi-infected HP24-treated cells.