Fig 2: A33 treatment increased Arg1-expressing microglia and myeloid-lineage cells at 3 hrs after TBI.Representative histogram overlay of iNOS+ and Arg1+ (A) infiltrating CD11b+ cells and (B) microglia at 3 hrs post-injury. Quantification of iNOS and Arg1-expressing (C) infiltrating CD11b+ cells and (D) microglia. Representative histogram overlay of iNOS+ and Arg1+ (E) infiltrating CD11b+ cells and (F) microglia at 24 hrs post-injury. Quantification of iNOS and Arg1-expressing (G) infiltrating CD11b+ cells and (H) microglia at 24 hrs after TBI. Mean ± SEM, n = 10/group (3 hrs), n = 5/group (24 hrs), *p<0.05, **p<0.01 vs. TBI+Vehicle, Student’s unpaired t-test. (iNOS = inducible nitric oxide synthase, Arg1 = arginase 1).

Fig 3: Inhibition of MAGL promotes the polarization of macrophages towards an M2 phenotype and alleviates pain behaviors in OA mice. (A-F) Immunohistochemical staining of MAGL, iNOS, and Arg1 in synovial tissues (upper panel, ×200, scale bar 50 µm; lower panel, ×400, scale bar 20 µm). (G) Representative images of the synovial tissues stained with H&E upper panel, ×200, scale bar 50 µm; lower panel, ×400, scale bar 20 µm). (H) Inflammation scores of the synovial tissues. (I) The mechanical pain threshold of mice was measured by electronic von Frey. (J) The thermal pain threshold of mice was measured using a hot plate (n=5 per group). *P<0.05, **P<0.01 vs. MIA-OA group; #P<0.05 vs. control group. ns, not significant; MAGL, monoacylglycerol lipase; OA, osteoarthritis; H&E, hematoxylin and eosin; MIA, monoiodoacetate.

Fig 4: CCR2 contributes to pathological type 1 inflammation during C. neoformans brain infection. (A and B) Frequencies of CD4+ T cells producing IFN-?, IL-17A, Foxp3, and IL-13 isolated from the brains of WT and CCR2-/- mice 21 dpi show a shift in the immune polarization away from the ultra-Th1 response toward a more-balanced, mixed Th-phenotype in CCR2-/- mice during CM. The pie charts show the ratio of different subsets of CD4+ T cells, Th1 (IFN-?+), Th2 (IL-13+), Th17 (IL-17A+), Tregs (Foxp3+), and other cells without detectable cytokine productions. (C) Total cell numbers of CD4+ T cells subsets isolated from the brains of WT and CCR2-/- mice at 21 dpi. (D and E) Expressions of iNOS and ARG1 by IM were analyzed in the brain of WT and CCR2-/- mice by intracellular flow cytometry. Note that IM produce a high level of iNOS in infected WT but shift to express ARG1 in infected CCR2-/- mice. (F and G) iNOS and ARG1 expression by microglia isolated from brains of WT and CCR2-/- mice with CM. Mean ± SEM from an experiment representative of two to four independent experiments (n > 4). *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Fig 5: CECs from anemic mice express ARG2 and have high levels of ROS.a Mean fluorescence intensity (MFI) of CellROX Green-FITC in CECs (CD71+TER119+) and RBCs (CD71-TER119+) of control (n = 6), NHA (n = 6), and HA-PHZ (n = 6) mice. Histograms show representative fluorescence of CellROX Green-FITC in CECs and RBCs from the spleens of the NHA mouse. P values were calculated with unpaired t-test. b MFI of CellROX Green-FITC in CECs (CD71+TER119+), CD45+ CECs (CD45+CD71+TER119+), leukocytes (CD45+TER119-), T cells (CD45+CD3e+), and myeloid cells (CD45+CD11b+) (n = 18). P values were calculated with Friedman’s test with Dunn’s post hoc test. c Percentages of ARG2+ CECs in control mice (n = 11), anemic mice (NHA, n = 5; HA-PHZ, n = 11; HA-TER119, n = 11), neonatal mice (n = 5), and isotype control-IgG-treated mice (control-IgG, n = 7) based on intracellular staining. d MFI of ARG2-APC in CECs from control mice, anemic mice (NHA, HA-PHZ, HA-TER119), neonatal mice, and isotype control-IgG-treated mice (each group n = 5). Histograms show the representative fluorescence of ARG2-APC in CECs in different groups and in anti-ARG2-unstained controls. e Percentages of ARG1+ CECs based on intracellular staining (n = 5). P values were calculated with one-way ANOVA with Dunnet’s post hoc test and with an unpaired t-test for HA-TER119. f Percentages of YFP+ CECs in reporter B6.129S4-Arg1tm1Lky/J mice (controls n = 4, NHA n = 8, HA-PHZ n = 8, neonatal n = 5, control-IgG n = 4, HA-TER119 n = 8). P values were calculated with one-way ANOVA with Dunnet’s post hoc test and with an unpaired t-test for HA-TER119. g MFI of YFP-FITC in CECs of reporter B6.129S4-Arg1tm1Lky/J mice in control mice (n = 4), anemic (NHA n = 8, HA-PHZ n = 4, HA-TER119 n = 8), neonatal mice (n = 5), and isotype control-IgG-treated mice (n = 4). P values were calculated with one-way ANOVA with Dunnet’s post hoc test. h Representative fluorescence of ARG1-YFP in CECs in reporter B6.129S4-Arg1tm1Lky/J control mice and anemic mice (NHA, HA-PHZ). Background fluorescence of YFP in CECs from wild-type C57Bl/6 mice presented as a negative YFP control. i, j Total arginase activity in CECs lysates (i n = 8) or in the supernatants from CECs cultures (j n = 8). P values were calculated with one-way ANOVA with unpaired t-test. k, l Percentages of ARG1+ CECs (k) isolated from the spleens of B6.129S4-Arg1tm1Lky/J incubated with diluent or PHZ (100 µM for 24 h) (n = 3) and representative fluorescence of ARG1-YFP (l). P values were calculated with unpaired t-test. Data show means ± SD. Each point in a–g, i–k represents data from individual mice. n values are the numbers of mice used to obtain the data. The source data underlying a–g, i–k are provided as a Supplementary Data 2 file.