Fig 5. LPS increases NO release from peritoneal macrophage and iNOS-derived NO has no effect on circadian transcription factor activity.

(A) NO release was measured by chemiluminescence detection of NO₂⁻ (mean ± SEM, n = 5, one-way ANOVA with Bonferroni post hoc correction, *p<0.05, versus control. # p<0.05, versus LPS). (B) Dual luciferase assay in COS cells expressing the Per1 promoter luciferase and BMAL1 and CLOCK or BMAL1, BMAL1+NPAS2+iNOS in the presence or absence of L-NAME (2mM), mean ± SEM, n = 6, one-way ANOVA with Bonferroni post hoc correction, *p<0.05, versus Bmal1 alone). (C) Peritoneal macrophages were synchronized as described and exposed to LPS (20ng/ml) with or without L-NAME (2mM). Bioluminescence was recorded every 2h for 48 hours. Oscillation curves were analyzed by cosinor and acrophase compared by one-way Anova with Bonferroni post hoc correction (mean ± SEM, n = 5, *p<0.05, versus Control, ns versus LPS).