Fig. 3: Neuroprotective effects of Withaferin A in PD are DJ1 dependent.

A Representative TH, GFAP, Iba1 and Nissl staining of SNc in WT and DJ1-KO mice, scale bar, 400 μm. B Unbiased stereological counts of TH⁺, GFAP⁺, Iba1⁺, and Nissl⁺ cells in SNc of WT and DJ1-KO mice. Data are mean ± s.e.m.; n = 9 biologically independent animals; *P < 0.05, **P < 0.01, and ***P < 0.001 by two-way ANOVAs followed by Tukey’s multiple comparisons test. C Representative photomicrographs of TH, GFAP, Iba1 staining in STR of WT and DJ1-KO mice, scale bar, 2 mm. D Stereological counts of TH, GFAP, Iba1 positive cells in STR of WT and DJ1-KO mice. Data are mean ± s.e.m.; n = 9 biologically independent animals; *P < 0.05, **P < 0.01, and ***P < 0.001 by two-way ANOVAs followed by Tukey’s multiple comparisons test. E Relative mRNA expression in SNc of WT and DJ1-KO mice. Data are mean ± s.e.m.; n = 8 biologically independent animals. F, G Representative immunoblots of TH, DAT, Nrf2 and STING in SNc (cropped blot images are shown, see Supplementary Fig. 16 for full immunoblots), quantification of TH, DAT, Nrf2, and STING levels. Data are mean ± s.e.m.; n = 9 biologically independent animals; *P < 0.05, **P < 0.01, and ***P < 0.001 by two-way ANOVAs followed by Tukey’s multiple comparisons test. H Time to traverse beam apparatus, time to descend pole, hind-limb clasping reflex score, fall latency from an accelerating rotarod and (I and J) gait analysis. Data are mean ± s.e.m.; n = 9 biologically independent animals. Two-way ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01, and ***P < 0.001. ns, not significant.