Extended Data Fig. 7: Mixed HCC fecal microbiota transplantation and fecal microbiota from two individual HCC patients impair gut barrier function and promotes bacteria translocation into liver in germ-free mice with or without DEN treatment.
(a) Gut permeability assay using FITC-dextran. n=6 (DEN+PBS), 12 (DEN+HD), 11 (DEN+HCC). (b) Representative pictures of Alcian blue staining, CLDN 3 and E-Cad IHC staining of colon tissues with quantitative analysis. n=6 biologically independent samples. (c) Representative bacterial culture of liver tissues under anaerobic and aerobic conditions with quantitative analysis. n=6 (DEN+PBS), 12 (DEN+HD), 11 (DEN+HCC). From left to right and top to bottom, the culture plates are blood agar plate, chocolate blood agar plate, MacConkey agar plate, and Columbia blood agar plate, respectively. (d) Images of Cy3-conjugated EUB338 probe detection. n=6 biologically independent samples. a–d are related to Extended Data Fig. 1a. (e) Pictures of CLDN 3 and E-Cad IHC staining, and Alcian blue staining of colon tissues with quantitative analysis. n=6 biologically independent samples. (f) Bacterial culture of liver tissues under anaerobic and aerobic conditions with quantitative analysis. n=10 (PBS), 12 (HD-FMT), 9 (HCC-FMT). From left to right and top to bottom, the culture plates are blood agar plate, chocolate blood agar plate, MacConkey agar plate, and Columbia blood agar plate, respectively. (g) Gut permeability assay using FITC-dextran and (h) Cy3-conjugated EUB338 probe detection of liver tissue. n=6 biologically independent samples. e-h are related to Extended Data Fig. 4a. (i) Pictures of Alcian blue staining, CLDN 3 and E-Cad IHC staining of colon tissues with quantitative analysis. n=6 biologically independent samples. (j) Gut permeability assay using FITC-dextran. n=9 (PBS), 10 (DEN+PBS), 12 (DEN+IHD), 9 (DEN+IHCC). (k) Bacterial culture of liver tissues under anaerobic and aerobic conditions with quantitative analysis. n=9 (PBS), 10 (DEN+PBS), 12 (DEN+IHD), 9 (DEN+IHCC). i-k are related to Extended Data Fig. 1e. Data (excluding liver with live bacteria) are presented as mean ± SEM. Each data point in bar plots represents one mouse. Liver with living bacteria was calculated using Fisher’s exact test. Unless otherwise stated, statistical significance was calculated using one-way ANOVA. Adjustments were made for multiple comparisons.
