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. 2022 Jan 6;12:781147. doi: 10.3389/fimmu.2021.781147

Figure 2.

Figure 2

Expression of cytokines, mucin, and tight junctions in patient-derived IEC. FACS analysis of isolated IEC. Cells were isolated by digestion and purified using a 20%/40% Percoll gradient. Purity was determined by flow cytometric analysis using antibodies against human CD45-PerCP and EpCam-Alexa488. Unstained controls were used to determine the negative population (A). Purified epithelial cells were immediately lysed, total RNA isolated and cDNA synthesized. Relative gene expression was calculated using the 2-ΔCT method, with β2-microglobulin as the housekeeping gene (B–E). Scatter plots with bar show mean ± SEM. Significance was determined using the Kruskal-Wallis test (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). Fiber-low (n=30); Fiber-high (n=13); DC AG (n=10); DC G, HSCR (n=12); DC G, non-HSCR (n=6). Not detectable samples: Fiber-low: TNF-α (n=2); IL-10 (n=6); TGF-β (n=1); Muc2 (n=2); ZO-1 (n=4); occludin (n=2). Fiber-high: IL-10 (n=4); ZO-1 (n=1). DC AG: IL-8 (n=1); TNF-α (n=1); IL-10 (n=3). DC G, HSCR: IL-8 (n=1); TNF-α (n=1); IL-10 (n=3); ZO-1 (n=2); occludin (n=2). DC G, non-HSCR: IL-10 (n=2); ZO-1 (n=1); occludin (n=1). DC, descending colon; AG, aganglionic; G, ganglionic.