Histological analysis of orthotopic HCC mouse tumors.

Hep_Cell tumors (day 21), Hep_Frag tumors (day 28) and iAST tumors (day 56) were stained for: A, morphology (H&E); B, tumor stroma (FAP); C, proliferation (Ki67); D, immune cells (CD45); E, macrophages (F4/80); F, Kupffer cells (Clec4f); G, CD4⁺ T cells (CD4); H, CD8⁺ T cells (CD8) and I, tumor vasculature (CD31). E,F, Dashed lines in the images of F4/80 and Clec4F stainings indicate the border between vital liver (VL) and tumor (T). I, CD31 signal quantification (n = 5) indicates significantly higher tumor vessel density in iAST compared to Hep tumors. In the scatter plot, each dot represents the average of 5 images per tumor. Differences between groups were tested for significance using the one-way ANOVA followed by Tukey multiple comparison test (***p<0.001).