Fig. 7

aOECs reversed neuroinflammation through crosstalk of JAK1/STAT1/3/6 signaling pathways and NF-κB/SOCS1/SOCS3 cascades to orchestrate IL-4-mediated M2 polarization of microglia. a Representative western blot band showed the levels of total protein (JAK1, STAT1, STAT3, and STAT6) and phosphorylated protein for the above-mentioned molecules in microglial cells under indicated stimulation for 24 h. b Quantitative analysis of the levels of phosphorylated protein (JAK1, STAT1, STAT3, and STAT6) in a, which were quantified and normalized to their non- phosphorylation. (n = 3/group). c Representative western blot band showed the levels of NF-κB, SOCS1, SOCS3, and phosphorylated NF-κB expression in microglial cells under the indicated conditions. β-actin served as a loading control. d Quantitative analysis of the levels of phosphorylated NF-κB, SOCS1, and SOCS3 (n = 3/group). All data are presented as means ± SEM. *P < 0.05, **p < 0.01, and ***p < 0.001. e Western blot analysis of M1 and M2 marker expression in microglial cells treated with the indicated treatments (several selective antagonists AZD1402 for IL-4Rα, AS1517499 for STAT6, and LY3009104 for JAK1). f The representative photographs of neurons co-cultured with microglia in the indicated conditions. Note that microglia pre-treated with several selective antagonists significantly inhibited the neuronal survival and neurite outgrowth. Scale bar = 100 μm. g Quantitative assessment of the number of Tuj-1⁺ cells co-cultured with microglia treated with antagonist as indicated, respectively. h Quantitative assessment of average length of neurite under indicated treatments. *p < 0.05, **p < 0.01, and ***p < 0.001 vs the corresponding controls