Fig 1: Colonic lamina propria (LP) cells isolated from control and antibiotic-treated WT and Casp1 KO mice and stained with (A) CD45 (Leukocytes); (B) Ly6G (neutrophils); (C) CD3 (CD3+ T cells); (D) CD4 (CD4+ T cells); (E) CD11c (dendritic cells); and (F) F4/80 (macrophages). Bars represent the percentage of the indicated cell population. Representative figure of two individual experiments. Data are mean (SEM). n = 4–5/group. *: p < 0.05.
Fig 2: Macrophage phenotyping in peritoneal tissue. Representative flow cytometry histograms in peritoneal tissue from control and TUG-891-treated apoE-knockout mice. F4/80 and CD11b+ (A), F4/80, iNOS; CD11b+; F4/80 (B), CD206+; CD11b+; F4/80 (C). Adjacent graphs represent mean ± SEM values, (* p < 0.05; n = 10–14 per group).
Fig 3: Macrophage phenotyping in atherosclerotic plaque. Validation of macrophages morphometry by RT-PCR analysis confirmed a trend toward reduction of M1 markers and no significant increase in M2 genes in the aorta of GW9508-treated apoE-/- mice, (A) (* p < 0.05; ** p < 0.005; n = 6–8 per group). Representative micrographs showing immunohistochemical staining of aortic roots from control and TUG-891-treated apoE-knockout mice. F4/80 (green), nitric oxide synthase 2 (iNOS)/arginase 1 (red) and 4',6-diamidino-2-phenylindole (DAPI) (blue), confirming the respective macrophage phenotype (white arrows). Adjacent graphs represent mean ± SEM values, (B) (* p < 0.05; n = 5–7 per group).
Fig 4: Representative RT-qPCR expression of genes associated with (A) inflammasome (Pycard, Aim2, Nlrp3, Nlrc4, Nlrp6, Il-1ß and Il-18); (B) toll-like receptors (TLR2, -4, -5 and -7); antimicrobial peptides (AMPs-Rentlß and Reg3?), the mucus layer component Muc2 and (C) macrophage M1/M2 signature (F4/80, Fitz1, Arg1, Il-6 and Il-12ß), the colon of WT and Casp1 KO antibiotic-treated groups. Data are mean (SEM). n= 7–9/group. *: p < 0.05, **: p <0.001. Dashed line indicates a background value of 1 of WT and Casp1 KO-control groups. Abx: Antibiotic.
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