Fig. 6

Treg modulates TMT-induced microglial activation and polarization. Time-lapse live imaging of BV2 microglial cells was monitored for 30 min with a CDr20 live microglia-specific probe (A). Representative images show microglial activity at 0 and 30 min with TMT treatment in the presence of CDr20 under the red fluorescent channel (excitation at 570 nm and emission at 600 nm). BV2 microglial cells were co-cultured with Tregs and stimulated with TMT for 24 h. The secreted levels of TNF-α and TGF-β (B) were measured using ELISA with supernatants. Relative mRNA expression of M1 cytokines, TNF-α and IL-1β; M1 microglial maker, NOS2; M2 cytokine TGFβ; M2 microglial makers, Mrc1 and Ym1 was examined and normalized to actin (C). Data are presented as the mean ± SEM. Significance was determined by Bonferroni’s correction and Tukey’s HSD (*p < 0.05, **p < 0.01, ***p < 0.001). Scale bar = 50 μm