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. 2022 Mar 21;13:847360. doi: 10.3389/fimmu.2022.847360

Figure 5.

Figure 5

AQP2 silencing directly inhibit astrocyte activation and indirectly reduce microglia migration. (A) Astrocyte were transfected with siAQP2 or siRNA-NC, and expression of AQP2, GFAP, TLR4, NF-kB and IL-1β were assessed by western blotting (n = 3/group). (B) Immunofluorescence of AQP2 (red signal) and GFAP (green signal) in astrocyte in astrocyte treated with siAQP2 or siRNA-NC. Astrocyte cell nuclei were immunostained (blue, DAPI). Scale bar: 10 μm. (C–E, G, H) Quantification of the relative protein expression levels of AQP2, GFAP, TLR4, NF-kB and IL-1β. (*p < 0.05 vs. siRNA-NC group; **p < 0.05 vs. siRNA-NC exposed to hemin; #p < 0.05 siRNA-NC group vs. siRNA-NC group exposed to hemin). The data are presented as the mean ± s.d. of three independent experiments. (F) Microglia were treated with medium of astrocyte for 24h transfected with siRNA or si-AQP2 RNA (with or without hemin). CD206, CD68 and CD86 were assessed by western blotting (n = 3/group). Quantification of the relative protein expression level of CD206 (I), CD68 (J) and CD86 (K) is shown. (*p < 0.05 vs. siRNA-NC group; **p < 0.05 vs. siRNA-NC exposed to hemin; #p < 0.05 siRNA-NC group vs. siRNA-NC group exposed to hemin). The data are presented as the mean ± s.d. of three independent experiments.