<i>Setd7</i> regulates IEC turnover responses after <i>T</i>. <i>muris</i> infection.

<p>(A) H&E stained caecal sections of <i>Setd7</i>^f/f and <i>Setd7</i>^ΔIEC mice at day 14 and day 21 post infection with <i>T</i>. <i>muris</i>. Yellow lines indicate crypt length. Original magnification is 100X. Bar = 50 μm (B) Crypt length (au = arbitrary units) of caecums of naïve (day 0) and <i>T</i>. <i>muris</i> infected mice (day 14 and day 21 post infection). (n≥30 of at least 4 mice, * P<0.05, *** P<0.001). (C) Proliferation as measured by counting Ki67+ cells per crypt from images such as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005876#ppat.1005876.s004" target="_blank">S4A</a>. (n≥20 of at least 4 mice, *** P<0.001). (D) IEC turnover was determined by dividing Ki67+ cells by total DAPI+ cells from images such as shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005876#ppat.1005876.s004" target="_blank">S4A</a>. Of note, crypt length correlated with total DAPI+ cells per crypt at all time points (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005876#ppat.1005876.s004" target="_blank">S4C</a>). (n≥20 of at least 4 mice, *** P<0.001).</p

Inhibition of YAP-TEAD interactions <i>in vivo</i> in resistance to <i>T</i>. <i>muris</i>.

<p>(A) IEC-specific expression of Wnt (<i>Lgr5</i>) and Hippo (<i>Ctgf</i>) target genes in <i>T</i>. <i>muris</i> infected <i>Setd7</i>^f/f (black bars) and <i>Setd7</i>^ΔIEC (grey bars) mice. Expression is relative to naïve mice (white bars). (n≥8, ** P<0.01). (B) Worm burdens at day 21 post infection of wild type (WT) mice. Mice were either vehicle treated (black bars) or vertepofin (VP) treated (striped bars) (n≥6). (C) Gene expression of <i>Il13</i> and <i>Ifng</i> from proximal colon at day 21 post infection. Expression is relative to uninfected, untreated WT mice (white bars, WT naive). (n≥6, * P<0.05) (D & E) IEC-specific expression of effector (<i>Tslp</i> and <i>Muc5AC</i>) and Hippo and Wnt target (<i>Ctgf</i> and <i>Lgr5)</i> genes. Expression is relative to naïve mice (white bars). (n≥6, *** P<0.001).</p

Deletion of <i>Setd7</i> specifically in IECs renders mice resistant to <i>T</i>. <i>muris</i>.

<p>(A) <i>Setd7</i>^f/f (black bars) and <i>Setd7</i>^ΔIEC (grey bars) mice were infected with 200 <i>T</i>. <i>muris</i> eggs and worm burdens were determined at day 14 or day 21 post infection (n≥9 for day 14, n = 6 for day 21, *** P<0.001). (B) Mesenteric lymph node cells from infected mice were re-stimulated for 72 h. IL-13 and IFN-γ concentrations in supernatants was determined by ELISA. (n≥4). (C) Expression of <i>Tslp</i> and <i>Muc2</i> in proximal colon at day 14 post infection with <i>T</i>. <i>muris</i> was assessed by qPCR. Expression is relative to infected control (<i>Setd7</i>^f/f) mice. (n≥8). (D) Protein levels of RELMβ that was secreted into the gut lumen of naïve and <i>T</i>. <i>muris</i> infected mice evaluated by Western blot.</p

<i>Setd7</i>^ΔIEC mice do not have increased resistance against infection with <i>H</i>. <i>p</i>. <i>bakeri</i>.

<p>(A) <i>Setd7</i>^f/f and <i>Setd7</i>^ΔIEC mice were infected with ~ 200 <i>H</i>. <i>p</i>. <i>bakeri</i> eggs and were killed day 28 post infection. Worm burdens were determined microscopically from the small intestine. (B) Mesenteric lymph node (mLN) cell counts from naïve and <i>H</i>. <i>p</i>. <i>bakeri</i> infected mice (day 28 post infection). (C) Gene expression of indicated genes in small intestine (SI) that was adjacent to infection site at day 28 post infection with <i>H</i>. <i>p</i>. <i>bakeri</i>. Expression is relative to naïve mice (white bars). (D) Periodic acid-Schiff (PAS) staining of small intestinal tissue sections of indicated mice at day 28 post infection. Bar = 100 μm. (E) PAS+ cells per villus were counted from images as shown in (D). (A-E) n = 3 (naives), n = 8 (infected) from 2 independent experiments. <i>Setd7</i>^f/f black bars, <i>Setd7</i>^ΔIEC grey bars.</p