Journal: Cell reports

Article Title: IL-22 promotes the formation of a MUC17 glycocalyx barrier in the postnatal small intestine during weaning.

doi: 10.1016/j.celrep.2021.108757

Figure Lengend Snippet: Figure 7. IL-22 primes postmitotic progenitor enterocytes to express Muc17 (A) Mice were administered rIl22 or PBS by i.p. injection at P14, and ileal tissues were collected for immunohistochemical analysis at P16. Ileal sections were stained for Muc17 (green), Ezrin (magenta), and DNA (gray). Scale bars 50 mm and 25 mm in corresponding insets, marked by roman numerals. (B) Mice were administered rIl22 and EdU at P14. Ileal tissues were collected for immunohistochemical analysis 2, 24, and 48 h post-injection. Ileal sections were stained for Muc17 (green), EdU (magenta), and DNA (gray). Boxed numbers indicate forerunner Muc17+ enterocyte position (yellow arrows) in relation to EdU+ IEC position (magenta arrows). Scale bars 50 mm and 25 mm in corresponding insets, marked by roman numerals. Data are represented as mean ± SD. (C) Confocal sections of P14 ileum stained for Il22ra1 (yellow) and DNA (gray). Yellow brackets indicate the intestinal crypt region. Scale bars 50 mm. See also Figure S6. (D) Mice were intraperitoneally injected with EdU at P16. Ileal tissues were collected for immunohistochemical analysis 2 , 24 (corresponding to P17), 48 (P18), 72 (P19), and 96 h (P20) post-injection. Ileal sections were stained for Muc17 (green), EdU (magenta), and DNA (gray). Arrows in magenta point to forerunner EdU+

Article Snippet: Primary antibodies were mouse anti-Actin C4 monoclonal antibody (1:10,000) (RRID: AB_2223041, #MAB1501, Millipore), mouse anti-Ezrin monoclonal antibody (1:500) (RRID: AB_476955, #E8897, Sigma-Aldrich), rat anti-Il22ra1 polyclonal antibody (1:100) (RRID: AB_1857199, #MAB42941, R&D Systems), rabbit anti-Lysozyme polyclonal antibody (1:500) (RRID: AB_10528813, #GWB-635D48, Genway Biotech), rabbit anti-Muc2 polyclonal antibody (1:500) (RRID: AB_1950958, #GTX100664, GeneTex Inc.), rabbit anti-Muc13 polyclonal antibody against human and mouse MUC13 (1:500) (Walsh et al., 2007), rabbit anti-MUC17C1 polyclonal antibody against human MUC17 (1:500) (Schneider et al., 2019), rabbit anti-Muc17S2 polyclonal antibody against mouse Muc17 (1:2,000) (Malmberg et al., 2006), mouse anti-Villin monoclonal antibody (1:50) (RRID: AB_2304475, #sc-58897, Santa Cruz).

Techniques: Injection, Immunohistochemical staining, Staining