Fig. 5

miR-124 attenuates LPS-induced inflammatory responses by targeting MEKK3/NF-κB signalling pathways in BV2 cells. The BV2 cells were transfected with miR-124 mimics or ctrl mimics. After 48 h, the production mRNA levels of MEKK3 (a) were determined using RT-qPCR, and western blot analysis was used to analyse the protein expression changes of MEKK3 (b) and p-p65 (c). NF-κB activity was analysed using luciferase assay (d). e, f The BV2 cells were transfected with miR-124 inhibitor or ctrl inhibitor, and the cells were harvested at 48 h. The relative expression of MEKK3 mRNA was evaluated using RT-qPCR (e), and the MEKK3 protein expression was studied using western blot analysis (f). g–j The BV2 cells were pre-transfected with MEKK3 siRNA for 48 h, and then the cells were transfected with miR-124 mimic or miR-124 inhibitor. After 24 h, cells were harvested, and the expression levels of p-p65 protein were evaluated using western blot analysis (g). RT-qPCR confirmed the expression of pro-inflammatory cytokines of TNF-α (h), iNOS (i), and IL-6 (j). The data were normalised against GAPDH. Data are normalised to saline controls and presented as the mean ± SD of three independent experiments. The fold change is statistically significant. *P < 0.05, **P < 0.01. NS not significant