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. 2020 Aug 24;223(8):1478–1487. doi: 10.1093/infdis/jiaa521

Figure 3.

Figure 3.

T5/N4/Rag-TKO mice display reduced barrier function, microbiota encroachment, and increased levels of neutrophils. A, Measure of serum FITC 3 hours following oral gavage with FITC-dextran. B and C, Assay of colon expression of indicated tight junction genes by qRT-PCR. D and E, Assessment of bacterial localization in Carnoy-fixed colon by confocal microscopy: (D) representative fluorescent image of bacteria in the colon and (E) average distance of closest bacteria per high-powered field. F, Fecal Lcn-2 measured by ELISA. G, Representative flow cytometry plot included neutrophils (CD45+CD11b+ MHCII-Ly6G+) and macrophages (CD45+CD11b+ MHCII+). H and I, Percentage of neutrophils (H) and macrophages (I) in colon. J and K, Relative level of MCP-1 (J) and TNF-α (K) in the colon analyzed by qRT-PCR. Data are means ± SEM, n = 3–4 mice per condition. Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; ELISA, enzyme-linked immunosorbent assay; FITC, fluorescein isothiocyanate; IEC, intestinal epithelial cell; Lcn2, lipocalin-2; MCP-1, monocyte chemoattractant protein-1; N4, Nod-like receptor 4;NS, nonsignificant; qRT-PCR, quantitative reverse transcription polymerase chain reaction; SSC, side scatter; T5, Toll-like receptor 5; TKO, triple knockout; TNF-α, tumor necrosis factor-α; ZO-1, zonula occludens-1. *P ≤ .05, **P ≤ .01 or ***P ≤ .001. n.s, not significant.