Fig. 8

MC1R activation on neuroinflammation via cAMP/PKA/Nurr1 signaling pathway at 48 h post-HI. a Representative picture of Western blot data showing bands of the expression levels of MC1R, cAMP, p-PKA, Nurr1, IL-1β, TNFα, and IL-6 either with BMS-470539 treatment alone, BMS-470539 + MC1R KO CRISPR, BMS-470539 + Nurr1 KO CRISPR, and BMS-470539 + control CRISPR groups. b–h Western blot data quantification of bands showed that BMS-470539 treatment significantly increased the expression of MC1R, cAMP, p-PKA, and Nurr1 compared to the HI + vehicle group. Knockout of MC1R using CRISPR significantly decreased MC1R, cAMP, p-PKA, and Nurr1 expression levels compared to the HI + BMS-470539 group or HI + BMS-470539 + control CRISPR group. Furthermore, knockout of Nurr1 using CRISPR significantly decreased the levels of Nurr1, but did not affect MC1R, cAMP, and p-PKA expression compared to the corresponding controls. Activation of MC1R with BMS-470539 showed significantly decreased levels of IL-1β, TNFα, and IL-6, while both treatment groups with CRISPR interventions significantly reversed these effects. Data was represented as mean ± SD. The one-way ANOVA was followed by Tukey’s post hoc test (*p < 0.05 versus sham; ^#p < 0.05 versus HI + vehicle; ^@p < 0.05 HI + BMS-470539 or HI + BMS-470539 + control CRISPR; n = 6 per group)