Additional file 1 of Akkermansia muciniphila and its membrane protein ameliorates intestinal inflammatory stress and promotes epithelial wound healing via CREBH and miR-143/145
Creators: name: Wade, Henry
name: Pan, Kaichao
name: Duan, Qihua
name: Kaluzny, Szczepan
name: Pandey, Ekta
name: Fatumoju, Linda
name: Saraswathi, Viswanathan
name: Wu, Rongxue
name: Harris, Edward N.
name: Su, Qiaozhu
Publisher: name: figshare
Year: 2024
Subjects: subject: Genetics
subject: FOS: Biological sciences

description: Additional file 1: Figure S1.Measurement of disease activity index over a 14-day period in DSS treated mice with or without A. muciniphila treatment.Immunohistochemistrystaining of anti-F4/80 antibody in the mouse distal colonic tissues as described in the Additional file 1: Supplemental Methods. Results represent the means±SD. The two-tailed Student’s t-test was used for statistical analyses of two-group comparisons. *P &lt; 0.05 and **P &lt; 0.01 versus controls.&gt;. Figure S2.qRT-PCR analysis of CREBH mRNA expression in the ileum of WT micepre-treatment with A. muciniphilaor Vehfollowed by DSStreatment for 14 days as described in the Methods.Body weights of the CREBH-KO micepre-treated with A. muciniphilaor Vehfollowed by DSStreatment for 14 days as described in the Methods. Results represent the means±SD. The two-tailed Student’s t-test was used for statistical analyses of two-group comparisons. *P &lt; 0.05 and **P &lt; 0.01 versus controls.&gt;. Figure S3.qRT-PCR analysis of mRNA expression of CLDN2 in Caco2 cells treated with TNFαfor 12 and 24 hrs, respectively.qRT-PCR analysis of mRNA expression of CLDN2 in Caco2 cells treated with IL-1βfor 24 and 48 hrs, respectively.qRT-PCR analysis of mRNA expression of CLDN5, CLDN8, OCLN, and ZO-1 in Caco2 cells transfected with mock vectoror vector expressing constitutively active CREBHfor 48 hrs.qRT-PCR analysis of mRNA expression of CLDN5, CLDN8, OCLN, and ZO-1 in Caco2 cells transfected mock or constitutively active CREBHfor 48 hrs and treated with or without TNFαfor 24 hrs. Results represent the means ± SD. The two-tailed Student’s t-test was used for statistical analyses of two-group comparisons. *P &lt; 0.05 and **P&lt;0.01 versus controls. &gt;. Figure S4.qRT-PCR analysis of mRNA expression of IGF1R in Caco2 cells transfected with mock vectoror vector expressing constitutively active CREBHfor 48 hrs.qRT-PCR analysis of mRNA expression of IGF1R in Caco2 cells transfected mock or constitutively active CREBHfor 24 h followed by additional 24 h treatment with TNFα.The non-outlined microscopy images of the wounded Caco2 cell monolayer shown in Figure 4F. Results represent the means ± SD. The two-tailed Student’s t-test was used for statistical analyses of two-group comparisons. **P &lt; 0.01 versus controls. &gt;. Figure S5.qRT-PCR analysis of mRNA expression of IGFBP1 and IGFBP4 in the colon tissues of the WT and CREBH-KO mice treated with DSSor vehiclein drinking water for 14 days.qRT-PCR analysis of mRNA expression of IGFBP1 and IGFBP4 in Caco2 cells transfected with mock vectoror vector expressing constitutively active CREBHfor 48 hrs.miR-143-3p and miR-203-3p response elements within the 3’- UTR of human, rat and mouse IGFBP5 mRNA predicted by TargetScan 7.2.Expression of miR-203-3p in the ileums of WT and CREBH-KO mice treated with either Vehor TNFα for 5 hrs.mRNA expression of IGFBP5 in Caco2 cells treated with mockor TNFαfor 24 hrs.qRT-PCR analysis of mRNA expression of IGFBP5 in Caco2 cells transfected with mock vectoror vector expressing constitutively active CREBHfor 48 h and treated with or without TNFαfor 24 hrs. Results represent the means with SD. The two-tailed Student’s t-test was used for statistical analyses of two-group comparisons. *P &lt; 0.05 versus controls. Figure S6.Immunoblot analysis of CLDN2 and β-actin in Caco2 cells transfected with mock vectoror vector expressing Amuc_1100 for 48 hrs.mRNA expression of CREBH, CLDN5, CLDN8, IGF1R, and IGFBP5 in Caco2 cells transfected with mock vectoror vector expressing constitutively active CREBHfor 48 h and treated with or without TNFαfor 24 hrs. Results represent the means with SD. The two-tailed Student’s t-test was used for statistical analyses of two group comparisons. **P &lt; 0.01 versus controls. &gt;. Table S1. Sequences of primers used in this study. Table S2. Scoring system for disease activity index.