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Figure 3: Western blotting for TNF-α and COX-2. ((a), (c)) Immunoblots of iNOS−/− control (Cont), cuprizone (CPZ), and cuprizone plus sildenafil (CPZ + Sild) groups. ((b), (d)) Graphs represent quantification and statistical analysis. The control group showed basal expression of TNF-α. CPZ treatment caused a significant increase of this cytokine, and sildenafil plus CPZ did not decrease its expression, which remained higher in relation to the control group. Only minimum amounts of COX-2 were present in the control group. CPZ and CPZ + Sild caused a significant increase of this enzyme, compared to control. The experiment was performed in triplicate ( animals/group). The results were expressed as mean ± SE. * , ** compared to control.