Fig. 5
miR-143/-145 induced by A. muciniphila target IGFBP5 in IGF signalling and enhances intestinal epithelial regeneration. Four groups of WT and CREBH-KO mice (n = 5–10/groups) were treated with either DSS or Veh (H2O) as described in "Methods". Colon and ileum tissues were collected for protein and RNA extractions for the following analysis. A Immunoblotting analysis of IGFBP5 and loading control of β-actin in the colon tissues of these mice. B Caco2 cells were transfected with mock (empty vector) or an active CREBH (N-terminal CREBH, constitutively active) plasmid for 48 h. Total RNAs were extracted for measurement of mRNA expression of IGFBP5 by qRT-PCR. C Expression of miR-143 and miR-145 in the colon tissue of DSS treated mice measured by qRT-PCR and normalized to small nucleolar RNA202. D Expression of miR-143 and miR-145 in the ileum tissues of CREBH-KO mice treated with either TNFα (60 ng/g body weight) or vehicle (Veh, saline) for 5 h (n = 5–6/group). E–G Four groups of the mice (n = 5–10/groups) were pre-treated with A. muciniphila (AKK) or Veh (PBS) followed by Veh (H2O) or DSS treatment as described in the "Methods". E Expression of miR-143 and miR-145 in the colon tissue measured by qRT-PCR and normalized to small nucleolar RNA202. F mRNA expression of IGFBP5 in the colon tissues of the treated mice. G Immunoblotting analysis of IGFBP5 and β-actin in ileum tissues of the indicated mice. Results represent the means ± SD. The two-tailed Student’s t-test was used for statistical analyses of two-group comparisons. *P < 0.05 and **P < 0.01 versus controls