Figure 6. Microglial activation was induced by neuron-derived CCL20 in vitro. (A) The effect of CCL20 siRNA on OxyHb-induced CCL20 expression was analyzed by Western blotting (n = 3 per group). (B) The effect of CCL20 on OxyHb-induced neuronal apoptosis was analyzed by TUNEL assay (n = 3 per group). (C–E) Cultured microglia were subjected to OxyHb-untreated conditioned medium (CM[-]), OxyHb-treated conditioned medium (CM[+]), and OxyHb CM from neurons pretreated with CCL20 siRNA for 72 h (CCL20 siRNA CM[+]) for 24 h. Western blotting (C) was used to determine CCR6 and Iba1 levels (n = 3). The IL-1β and TNF-α expression levels were analyzed by RT-qPCR (D) and ELISA (E) (n = 3 per group). **P<0.01. (F–H) Cultured microglia pretreated with NC and CCR6 siRNA for 72 h were exposed to CM[+]. Then, Western blotting (F) was used to determine CCR6 and Iba1 levels (n = 3). RT-qPCR (G) and ELISA (H) were used to determine IL-1β and TNF-α levels (n = 3 per group). Data were presented as means ± SD. **P < 0.01.