Extended Data Fig. 10: TLR4 inhibition reverses HCC-FMT induced-HCC development.
(a) Design of TLR4i on HCC-FMT induced HCC promotion in germ-free mice with DEN treatment (GFD/PBS group n=8; GFD/HCC-FMT group n=15; GFD/HCC-FMTi group n=10) and relevant results (b–g). TLR4i was daily gavaged in this experiment. (b) Representative images of liver gross morphology (yellow circles indicate tumor area), and H&E staining of mice liver sections with tumor incidence, tumor number, and tumor burden. n=8 (GFD/PBS), 15 (GFD/HCC-FMT), 10 (GFD/HCC-FMTi). (c) Representative bacterial culture of liver tissues under anaerobic and aerobic conditions with quantitative analysis. n=8 (GFD/PBS), 15 (GFD/HCC-FMT), 10 (GFD/HCC-FMTi). 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. Gut permeability assay using 500 kDa FITC-dextran. n=6 biologically independent samples. (d) Representative pictures of Alcian blue staining, E-Cad IHC staining, and Cy3-conjugated EUB338 probe FISH detection of colon tissues with quantitative analysis. n=6 biologically independent samples. (e) Cy3-conjugated EUB338 probe FISH, Cy3-conjugated K. pneumoniae probe FISH, TLR4, PCNA, Ki-67 IHC of liver sections. n=6 biologically independent samples. (f) α-SMA, Desmin IHC staining, and Sirius red staining of liver sections. n=6 biologically independent samples. (g) Mouse Cancer Pathway Finder Array and Inflammatory Response and Autoimmunity PCR Array of liver tissues. n=3 independent experiments with similar results. Data (excluding tumor incidence, liver with live bacteria and PCR array results) are presented as mean ± SEM. Each data point in bar plots represents one mouse. Tumor incidence and Liver with live bacteria were calculated using Fisher’s exact test. Unless otherwise stated, statistical significance was calculated using one-way ANOVA. Adjustments were made for multiple comparisons.
