Fig. 3. P35 expression in RAW 264.7 cells does not inhibit iNOS expression and nitrite production. RAW 264.7 cells or vector- or p35-transfected clone (1x10⁶ cells/well) in 6-well tissue culture plates were treated with cisplatin (2 µg/ml) and IFN- (25 U/ml), or LPS (1 µg/ml) and IFN- (25 U/ml) in the presence or absence of L-NMMA (1 mM) as before. Cells were also treated directly with NO donor, SNP (1 mM). After 18 hours of treatment, cell extracts were examined for -iNOS expression by immunoblotting (A). Culture supernatants were assayed for nitrite after 18 hours of culture as described in Materials and Methods (B). Data shown are means±s.e.m. and are representative of three independent experiments done in triplicate. ^*P<0.05 versus values of control cultures. ^**P<0.05 versus values of cisplatin and IFN-/LPS and IFN--treated cultures.