Journal: Cell reports
Article Title: Mucolytic bacteria license pathobionts to acquire host-derived nutrients during dietary nutrient restriction.
doi: 10.1016/j.celrep.2022.111093
Figure Lengend Snippet: Figure 4. Disruption of colonic mucus barrier under L-serine starvation enhances the encroachment of AIEC to the epithelial niche (A) Relative abundances of A. muciniphila and E. coli during DSS treatment were each assessed by qPCR. (B and C) Colonic sections were stained with Alcian blue/periodic acid-Schiff (AB/PAS), and the thickness of the inner mucus layer was measured (scale bars, 100 mm). (D) Intestinal permeability was assessed with fluorescein isothiocyanate (FITC)-dextran. (E) Immunostaining (MUC2, green; DAPI, blue) and fluorescence in situ hybridization (FISH) (EUB338 probe, red) of Carnoy’s-solution-fixed colonic sections (scale bars, 100 mm). (F) SPF C57BL/6 mice were fed the Ctrl diet or the DSer diet for 3 days and then given 1.5% DSS for 5 days, followed by conventional water for 2 days. Mice were infected with each strain of E. coli (1 3 109 colony-forming units [CFUs]/mouse) on days 5 and 6. On day 7 post-DSS, all mice were euthanized. (G) Homogenates of colon tissues were cultured on Luria-Bertani (LB) agar plates supplemented with ampicillin or streptomycin. The number of viable bacteria was estimated by counting the CFUs and calculating the fold change (DSer diet and Ctrl diet). Data are representative of two independent experiments (n = 4–5). Dots indicate individual mice, with mean ± SEM or geographic mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001 by one-way ANOVA with Tukey post hoc test or unpaired t test.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Alexa Fluor 488 goat anti-rabbit IgG Invitrogen Cat# A11008; RRID: AB_143165 Alexa Fluor 555 goat anti-mouse IgG Invitrogen Cat# A32727; RRID: AB_2633276 Alexa Fluor 555-conjugated EUB338 probe Invitrogen N/A Anti-E. coli LPS antibody Abcam Cat# ab35654; RRID: AB_732222 Mucin 2 antibody (H-300) Santa Cruz Biotechnology Cat# sc-15334; RRID: AB_2146667 Bacterial and virus strains Adherent-invasive Escherichia coli: LF82 strain Darfeuille-Michaud et al., 1998 N/A Adherent-invasive Escherichia coli: LF82 DTS mutant strain Kitamoto et al., 2020 N/A Adherent-invasive Escherichia coli: LF82 DfimH mutant strain Imai et al., 2019 N/A Adherent-invasive Escherichia coli: CUMT8 strain Kitamoto et al., 2020 N/A Akkermansia muciniphila: DSM 22959, type strain DMSZ DMS 22959 Bacteroides ovatus: DSM 1896, type strain DMSZ DSM 1896 Bacteroides uniformis: ATCC 8492, type strain ATCC ATCC 8492 Clostridium symbiosum: DSM 934, type strain DMSZ DSM 934 Collinsella aerofaciens: DSM 3979, type strain DSMZ DSM 3979 Desulfovibrio piger: ATC 29098, type strain ATCC ATCC 29098 Escherichia coli: HS strain ATCC N/A Escherichia coli: MG1655 strain Miranda et al., 2004 N/A Eubacterium rectale: DSM 17629, A1-86 DSMZ DSM 17629 Faecalibacterium prausnitzii: DSM 17677, A2-165 DSMZ DSM 17677 Marvinbryantia formatexigens: DSM 14469, type strain, l-52 DSMZ DSM 14469 Roseburia intestinalis: DSM 14610 type strain, L1-82 DSMZ DSM 14610 Chemicals, peptides, and recombinant proteins Acetic acid Thermo Fisher Scientific A38S Alcian blue Sigma-Aldrich A5268 Ampicillin Sandoz Inc 0781-3408 BHI agar BD Difco BD241830 Bovine Serum Albumins Thermo Fisher Scientific BP1600 Chloroform Thermo Fisher Scientific 423550250 DAPI Sigma-Aldrich D9542 Dextran Sodium Sulfate MP Biomedical 0216011090 DMEM Gibco 11320033 Ethanol Thermo Fisher Scientific BP28184 FITC-dextran Sigma-Aldrich FD4 HBSS Thermo Fisher Scientific 14170112 (Continued on next page) e1 Cell Reports 40, 111093, July 19, 2022
Techniques: Disruption, Staining, Permeability, Immunostaining, In Situ Hybridization, Infection, Cell Culture, Bacteria
ATCC is a verified supplier 
ATCC manufactures this product 
![Figure 4. Disruption of colonic mucus barrier under L-serine starvation enhances the encroachment of AIEC to the epithelial niche (A) Relative abundances of A. muciniphila and E. coli during DSS treatment were each assessed by qPCR. (B and C) Colonic sections were stained with Alcian blue/periodic acid-Schiff (AB/PAS), and the thickness of the inner mucus layer was measured (scale bars, 100 mm). (D) Intestinal permeability was assessed with fluorescein isothiocyanate (FITC)-dextran. (E) Immunostaining (MUC2, green; DAPI, blue) and fluorescence in situ hybridization (FISH) (EUB338 probe, red) of Carnoy’s-solution-fixed colonic sections (scale bars, 100 mm). (F) SPF C57BL/6 mice were fed the Ctrl diet or the DSer diet for 3 days and then given 1.5% DSS for 5 days, followed by conventional water for 2 days. Mice were infected with each strain of E. coli (1 3 109 colony-forming units [CFUs]/mouse) on days 5 and 6. On day 7 post-DSS, all mice were euthanized. (G) Homogenates of colon tissues were cultured on Luria-Bertani (LB) agar plates supplemented with ampicillin or streptomycin. The number of viable bacteria was estimated by counting the CFUs and calculating the fold change (DSer diet and Ctrl diet). Data are representative of two independent experiments (n = 4–5). Dots indicate individual mice, with mean ± SEM or geographic mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001 by one-way ANOVA with Tukey post hoc test or unpaired t test.](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_8565/pm35858565/pm35858565__page7_image1.jpg)
