Figure 6. JAML regulates macrophage phenotypic polarization via a Mincle-dependent mechanism.

(A and B) BMDMs from WT or Jaml^–/– mice were serum starved for 4 hours (M0) and then polarized to M1 (LPS) or M2 (IL-4) for 8 hours. Media were removed, M1 macrophages were treated with M2 stimuli (IL-4), and M2 macrophages were treated with M1 stimuli (LPS) for an additional 8 hours. Real-time PCR was performed to measure Arg1 and Ccl8 expression in M1 macrophages polarized to M2 (A). Il6 and iNos gene expression was measured in M2 macrophages polarized to M1 (B) (n = 7). (C) Flow cytometry analysis of renal macrophages in the injured kidney after IRI. CD45-positive cells were divided into the F4/80^lo and F4/80^hi groups. Representative flow cytometry analysis of M1 (CD80^hi) and M2 (CD206^hi) cell populations in F4/80^lo and F4/80^hi macrophages isolated from the kidney in different groups of mice. SSC, side scatter. (D) Representative flow cytometry histogram showing cell surface marker CD206 (M2) and CD80 (M1) expression on 2 subtypes of macrophages and quantitative analysis of MFI of CD206-AF647 or CD80-PE-Cy7 (n = 6). Data are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA test (A and B), 2-tailed Student’s unpaired t test (D).