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. 2023 Mar 15;18(11):2429–2435. doi: 10.4103/1673-5374.371367

Figure 6.

Figure 6

Effect of LZ-3 on microglia polarization in vivo and in vitro.

(A) Representative images of immunofluorescence staining for Iba-1 (green, Alexa Fluor® 488) and iNOS (red, Alexa Fluor® 647) in microglia after LZ-3 treatment. LZ-3 reduced the number of Iba-1- and iNOS-positive cells compared with Mod. Arrows indicate double-stained microglia. (B) The average number of Iba-1+ and iNOS+ cells detected as shown in A (n = 3). (C) Representative images of immunofluorescence staining for Iba-1 (green, Alexa Fluor® 488) and CD206 (red, Alexa Fluor® 647) in microglia after LZ-3 treatment. LZ-3 increased the number of Iba-1- and CD206-positive cells compared with Mod. Arrows indicate double-stained microglia. Scale bars: 100 μm. (D) The average number of Iba-1+ and CD206+ cells detected as shown in C (n = 3). ##P < 0.01, ###P < 0.001, vs. sham group; **P < 0.01, vs. model group. (E, F) LZ-3, upadacitinib, or PBS was added to the BV2 cell culture medium after 24 hours of re-oxygenation after OGD. Relative mRNA levels of CD11b, CD32, and iNOS as detected by qPCR (E), and relative mRNA levels of Arg-1, IL-10, and CD206 (F) (n = 3). ##P < 0.01, vs. control group; *P < 0.05, **P < 0.01, vs. OGD group. Data are expressed as mean ± SD and were analyzed by one-way analysis of variance followed by Tukey’s post hoc test. Arg-1: Arginase 1; Con: control group; DAPI: 4’,6-diamidino-2-phenylindole; IL-10: interleukin-10; iNOS: inducible nitric oxide synthase; Mod: model group; OGD: oxygen-glucose deprivation.