Epithelial differentiation reduces inflammatory cytokine expression in colonic organoids.

<p>(A) RNA markers of stem cells (<i>Lgr5</i>), enterocytes (<i>Alpi</i>), and goblet cells (<i>Muc2</i>) and enteroendocrine cells <i>(Chga</i>) were measured by quantitative polymerase chain reaction (qPCR) and normalized to standard culture condition organoid expression levels. Conditions for enrichment of stem cells or specific differentiated lineages can be found in the Materials and methods section. Horizontal and vertical lines represent mean +/− standard error. Significance was determined by ANOVA with Tukey-Kramer post-test. * represents <i>p</i> ≤ 0.05; *** represents <i>p</i> ≤ 0.001. (B) Representative cytokines, chemokines, and growth factors were measured by Luminex and represented in pg/ml. Horizontal and vertical lines represent mean +/− standard error. Significance was determined by ANOVA with Tukey-Kramer post-test. * represents <i>p</i> ≤ 0.05; ** represents <i>p</i> ≤ 0.01. (C) A partial least squares regression (PLSR) model built on cytokine, chemokine, and growth factor measurements from the mouse experiment described in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2002417#pbio.2002417.g001" target="_blank">Fig 1</a> with organoid protein expression data projected. Differentiated organoids project with focal inflammation and control groups, while proliferative, stem-like organoids are projected with the continuous inflammation group. Original samples used to generate the model project into the gray rectangle (control), blue rectangle (focal inflammation), and red rectangle (continuous inflammation). Underlying numerical values for panels A–C are provided in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2002417#pbio.2002417.s001" target="_blank">S1 Data</a>. MCP-1, monocyte chemoattractant protein 1; MIG, mitokine induced by gamma interferon; MIP-1α, macrophage inflammatory protein 1-alpha.