Fig. 6

Knockdown of miR-195 induced microglial polarized toward M1 phenotype-dependent on the CX3CL1-CX3CR1 signaling pathway in vitro. a Schematic diagram of the neuron-microglia cell co-culture system. Neurons were transfected with miR-195, AMO-195, miR-195 + AMO-195, Cx3cl1-ODN, or NC for 48 h. Then, BV2 cells were seeded in the top compartment of the transwell with the NRNs were cultured in the bottom compartment. Subsequently, BV2 cells were co-cultured with NRNs for 24 h. b, c The effects of miR-195 on endogenous CX3CL1 expression in NRNs by western blotting (b) and immunofluorescence staining (c) after the NRNs were transfected with miR-195, AMO-195, miR-195 + AMO-195, miR-195+ Cx3cl1-ODN, or NC. d MiR-195 downregulated CX3CL1 expression in co-cultured BV2 cells assessed by immunofluorescence staining and western blotting. e MiR-195 did not affect CX3CR1 expression in co-cultured BV2 cells evaluated assessed by immunofluorescence staining and western blotting. f Downregulating miR-195 increased the ratio of CD68/CD206 in Iba-1+ cells in co-cultured BV2 cells. Bars represent the mean ± SD, n = 3 batches of cell culture. *P < 0.05 vs NC; ^#P < 0.05 vs AMO-195. Scale bar: 40 μm. All data were analyzed using one-way ANOVA followed by Tukey test