Fig. 7: The T cell immune activation and anti-tumor efficacy of AQ were evaluated in a humanized mouse model of hepatocellular carcinoma (HCC) and a patient-derived xenograft (PDX) model of HCC.

a Schematic of the humanized CDX model development and AQ therapy experimental protocols by figdraw.com. b IHC staining and semi-quantitative scoring of β-catenin; experiments were replicated three times with consistent results. c LC–MS-based targeted protein analysis of the Wnt/β-catenin pathway in AQ-treated and mock-treated tumors. d GSEA analysis of Wnt/β-catenin pathway data from AQ-treated and mock-treated tumors; p value calculated by permutation test. e Multichannel flow cytometry was used to analyze Treg and CTL populations in tumors from two treatment groups (n = 3 biological replicates per group). CTLs were identified as CD45 + CD8 + CD3+ cells, with GzmB+ or IFNγ + CTLs further characterized. Tregs were identified as CD45+ CD4+ CD25+ cells, with FOXP3+ Tregs further characterized. f ssGSEA of the proteomics data about Treg cells and CTLs. g, h Tumor photos (g) and tumor weights (h) from tumors. i The tumor growth curve upon treatment with AQ or a control substance (n = 5/group). j Schematic illustrating the development of a humanized PDX model and the experimental protocols for AQ and Anti-PD1 combo-therapy in this model by figdraw.com. k RNA-seq analysis of Wnt/β-catenin pathway in the four groups. l ssGSEA of tumor samples from four groups about Wnt/β-catenin pathway, cell cycle arrest and apoptosis (n = 3/group). m ssGSEA of tumor samples from four groups about activated CD8 T cell, CD8 TCR, and Tregs. (n = 3/group). n The tumor photos depict tumors treated with mock-treated, AQ-treated, anti-PD1-treated, and AQ/Anti-PD1 combination therapy. o The tumor growth curve is plotted for the treatment groups (n = 5/group). All box-whisker plots B&H center on the median; the bounds of the boxes mark the upper and lower quartile; the whiskers extend to the upper and lower extremes (1.5x interquartile range from the upper and lower quartiles). The error bars in e, i, l, m are presented as means ± SD. The p < 0.001 in b, h, i represents p = 10E-15. p value in b, e, h, i calculated by two-tailed t-test.