Figure 4.

15d-PGJ₂ inhibits production of iNOS protein and mRNA in LPS-stimulated BV-2 cells. (A) Cells were incubated with 80 ng/ml of LPS alone, LPS + 0.25, 0.5, 1, or 1.5 μM 15d-PGJ₂, 15d-PGJ₂ alone, or control buffer. Cell lysates were prepared at 9 h after treatment, and iNOS protein levels were determined by Western blotting. (B) Cells were incubated with 80 ng/ml of LPS alone, LPS + 2, 1, or 0.1 μM 15d-PGJ₂, the control buffer, or 2 μM 15d-PGJ₂ and total RNA isolated at 12h. One representative slot blot is shown. Levels of iNOS mRNA were normalized to glyceraldehyde-3-phosphate dehydrogenase levels and expressed as relative increase compared with the control. Values correspond to the mean ± SEM of four independent experiments.