mouse il-22 r alpha 1 antibody  (Bio-Techne corporation)

Journal: Nutrients

Article Title: Promoting Effect of L-Fucose on the Regeneration of Intestinal Stem Cells through AHR/IL-22 Pathway of Intestinal Lamina Propria Monocytes.

doi: 10.3390/nu14224789

Figure Lengend Snippet: Figure 2. Increased ISC regeneration and IL-22 secretion in mice treated with L-fucose. (A) Rep- resentative images of Ki67 immunohistochemistry of the colon (scale bar, 100 µm). (B) Ki67+ cell percentage in crypts. (C) Representative images of Lgr5 (green) in the colon (scale bar, 50 µm). (D) Lgr5-positive cell percentage in crypts. (E) Representative images of IL-22R (red) in the colon (scale bar, 50 µm). (F) IL-22R-positive cell percentage in crypts. Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) analysis of (G) Lgr5, Olfm4, Ascl2 and (H) IL-22 in the colon. (I) Western blotting analysis of Lgr5 and IL-22 proteins in the colon. (J) ELISA analysis of IL-22 in the colon. The scale bar represents 100 µm. Data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS, no significance.

Article Snippet: For immunofluorescent staining, after the same deparaffinization and antigen retrieval steps mentioned above, the slides were permeabilized with 0.3% Triton X-100 at room temperature for 20 min, followed by blocking with 10% donkey serum for 1 h. Then, the sections were incubated with anti-mouse Lgr5 antibody (1:200, A10545, ABclonal, Woburn, MA, USA) and anti-mouse IL-22R antibody (1:50, MAB42941, R&D system) at 4 ◦C overnight.

Techniques: Immunohistochemistry, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Nutrients

Article Title: Promoting Effect of L-Fucose on the Regeneration of Intestinal Stem Cells through AHR/IL-22 Pathway of Intestinal Lamina Propria Monocytes.

doi: 10.3390/nu14224789

Figure Lengend Snippet: Figure 7. IL-22R-p-STAT3 pathway mediated by IL-22 secretion of LPMCs in ISC proliferation after L-Fucose treatment. Representative immunofluorescence images of (A) IL-22R (red), Lgr5 (green) and (B) p-STAT3 (green) in organoids (scale bar, 100 µm). Quantitative analysis of (C) Lgr5, (D) IL-22R and (E) p-STAT3 fluorescence intensity in organoids. (F) Western blotting analysis of Lgr5, β-catenin, p-STAT3 and STAT3 in organoids. (G) qRT-PCR analysis of IL-22R in organoids. (H) ELISA analysis of IL-22 in the culture supernatant of the LPMC -organoid coculture system. Data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, **** p < 0.0001; NS, no significance. +, with. −, without.

Article Snippet: For immunofluorescent staining, after the same deparaffinization and antigen retrieval steps mentioned above, the slides were permeabilized with 0.3% Triton X-100 at room temperature for 20 min, followed by blocking with 10% donkey serum for 1 h. Then, the sections were incubated with anti-mouse Lgr5 antibody (1:200, A10545, ABclonal, Woburn, MA, USA) and anti-mouse IL-22R antibody (1:50, MAB42941, R&D system) at 4 ◦C overnight.

Techniques: Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Nutrients

Article Title: Promoting Effect of L-Fucose on the Regeneration of Intestinal Stem Cells through AHR/IL-22 Pathway of Intestinal Lamina Propria Monocytes.

doi: 10.3390/nu14224789

Figure Lengend Snippet: Figure 8. Restoration of TNF-α-induced ISC damage through IL-22 after L-fucose treatment. (A) Representative images of organoids under a light microscope (scale bar, 100 µm). (B) Rep- resentative images of EdU (red) staining of organoids (scale bar, 100 µm). (C) Representative images of IL-22R (red) immunofluorescence in organoids (scale bar, 100 µm). (D) Number of organoids per field of view. (E) Number of organoids with more than 3 buds per graph. (F) Average area of organoids per graph. Quantitative analysis of (G) EdU and (H) IL-22R fluorescence intensity in organoids. (I) ELISA analysis of TNF-α in the culture supernatant of the LPMC-organoid coculture system. (J) Western blotting analysis of Lgr5 protein in organoids. (K) qRT-PCR analysis of Lgr5 and Olfm4 in organoids. Data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS, no significance. +, with. −, without.

Article Snippet: For immunofluorescent staining, after the same deparaffinization and antigen retrieval steps mentioned above, the slides were permeabilized with 0.3% Triton X-100 at room temperature for 20 min, followed by blocking with 10% donkey serum for 1 h. Then, the sections were incubated with anti-mouse Lgr5 antibody (1:200, A10545, ABclonal, Woburn, MA, USA) and anti-mouse IL-22R antibody (1:50, MAB42941, R&D system) at 4 ◦C overnight.

Techniques: Light Microscopy, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

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