Fig. 7

Increasing expression levels of MEKK3, p-p65, and activated microglia are evidenced in the SNpc of MPTP-treated mice in vivo. The mice received one intraperitoneal injection of MPTP-HCl per day for five consecutive days, whereas the control mice received saline injections. Then, the mice were decapitated, and the midbrains were harvested at different time points after MPTP intoxication as follows: 0 (immediately after the last MPTP injection), 1, 7, and 21 days after the last MPTP injection. a The miR-124 expression level was determined using RT-qPCR and normalised with U6 RNA. The production mRNA level of MEKK3 (b) was determined using RT-qPCR. Western blot analysis evaluated the MEKK3 expression (c) in the total protein samples extracted from the midbrain. The data were normalised against GAPDH. d, e Immunohistochemical analysis was performed to analyse the MEKK3 response (d). Graphical representation of the MEKK3 IOD value after MPTP injection is shown (e). The scale bar represents 50 μm. f A confocal image supported by immunofluorescence confirmed the expression levels of MEKK3 and Iba1⁺. Green: anti-MEKK3; red: anti-Iba1 (antibody microglia). The scale bar represents 100 μm. g Western blot analysis determined p-p65 protein expression after MPTP injection. Data are normalised to those of saline controls and presented as the mean ± SD of three independent experiments. The fold change is statistically significant. *P < 0.05, **P < 0.01, ***P < 0.001