Fig. 4

Soluble factors from mouse brain glial cells induce M2 polarization of BMDM. a BMDM from Cx3cr1 ^+/GFP mice were cultured with or without primary glial cells from C57BL/6 mice in a 1:1 ratio for 1 or 3 days. Then the expression of CD206 (a M2 marker) was analyzed using flow cytometry. Representative histograms of CD206 expression in GFP⁺/CD45⁺ populations are shown. b The percentages of CD206⁺ populations in BMDM were calculated. Mean ± SEM of 3 independent experiments are shown (*p < 0.05). c BMDM were incubated in primary glia-conditioned media for 1 and 3 days. Then, the cells were harvested and stained with CD45-PE, CD11b-FITC, and CD206-APC for flow cytometry analysis. The percentages of CD206⁺ cells in CD45⁺/CD11b⁺ populations are analyzed. Mean ± SEM of 3 independent experiments are shown (*p < 0.05, ***p < 0.001). d–g Total RNA was isolated from BMDM at 1 or 3 days after glia-conditioned media treatment. Arg-1, Ym1, iNOS, and TNF-α mRNA expressions were measured using real-time RT-PCR. Mean ± SEM of at least 3 independent experiments are shown (*p < 0.05)