Fig 2: Gut microbiota regulates AML via alteration of intestinal barrier function and LPS blood concentration.Gut microbiota disorders in AML lead to a decrease in intestinal butyrate. Decreased butyrate weakens gut barrier function by altering tight junction proteins (claudin-1, claudin-2, ZO-1). Dysregulation of intestinal barrier function allows LPS leakage into blood, which in turn promotes the proliferation of AML cells and accelerates AML progression. These effects could be reversed by treatment with butyrate or FMT.

Fig 3: Ripk2 over-expression alleviates intestinal damage caused by Card9 deficiency in sepsis.WT mice and Card9-/- mice (n = 10 for each group) were treated with LPS after 24 h Ad-Ripk2 injected into tail vein. A Secretion of IL-1ß and IL-10 in serum was assessed by ELISA. B Production of AST and ALT was detected in serum. C Histological analysis of intestinal was shown by H&E staining. D Intestinal tissue from Card9-/- mice was staining with alcian blue. E Immumohistochemical analysis of ZO-1, Occludin and Claudin-2 in intestinal tissue from Card9-/- mice. Data are shown as mean ± SEM. Each group was representative of at least three biological replicates. **p < 0.01, ***p < 0.001.

Fig 4: Card9-/- mice are susceptible to sepsis and along with intestinal inflammation and gut barrier dysfunction.WT mice and Card9-/- littermates (n = 10 for each group) were intraperitoneal injection with LPS (5 mg/kg). A Spleen photo of mice in control and experimental groups. B Survival rate of WT mice and Card9-/- mice within 5 days after LPS treatment. C Secretion of IL-1ß, IL-10, IL-6 and TNF-a in serum was assessed by ELISA. D Histological analysis of intestinal was assessed by H&E staining. E Intestinal tissue from WT mice and Card9-/- mice were staining with alcian blue. F Western blot analysis of ZO-1, Occludin and Claudin-2 in intestinal tissue from WT-sepsis mice and Card9-/--sepsis mice. G Immunofluorescence analysis of ZO-1, Occludin and Claudin-2 in the intestinal mucosa. Data are shown as mean ± SEM. Scale bars, 0.1 mm. Each group was representative of at least three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 5: Cldn2 and Cldn1 mRNA expression after miR-195-5p mimic transfection and TNF-a stimulation. (A) Cldn2 mRNA expression was analyzed by Real-Time PCR after transfection with miR-195-5p mimic in TNF-a-stimulated HT-29, Caco2 and T84 human cell lines. The increase of intracellular amount of miR-195-5p (at 30 nM and 50 nM concentrations) led to a significant decrease of Cldn2 in all cell lines. (B) Cldn1 mRNA expression was analyzed by Real-Time PCR after transfection with miR-195-5p mimic in HT-29, Caco2 and T84 human cell lines. Cldn1 mRNA levels were slightly modulated after transfection but not in a significant manner except for T84 cell line. Mock indicates mock-transfected cells going through the transfection processes without addition of mimic miRNA. Expression data were normalized on the housekeeping gene Gapdh. Data are representative of four independent experiments (mean ± SEM). ** p < 0.001; *** p < 0.0001.