Fig. 5

The dysadherin–YAP axis modulates drug resistance and PD-L1–mediated immune evasion in HCC. a GSEA of DEGs between dysadherin^high and dysadherin^low tumors (GSE9843), revealing enrichment of immunotherapy resistance and drug resistance signatures. b Heatmaps showing increased expression of gene sets associated with resistance to tyrosine-kinase inhibitors (left), cytostatic drugs (middle), and doxorubicin (right) in dysadherin^high HCC tumors. c Cell viability and IC₅₀ curves of PLC/PRF/5 (dysadherin OE) and SK-Hep1 (dysadherin KD) cells treated with verteporfin or YAP5SA, in the presence of sorafenib. d Apoptosis analysis using Annexin V/PI staining in PLC/PRF/5 and SK-Hep1 cells under the same conditions as in (c). e Heatmap showing elevated expression of immunosuppressive genes and immune checkpoint–related transcripts in dysadherin^high tumors from GSE9843. f Immunoblot analysis of active YAP, phospho-YAP (S127), total YAP, and PD-L1 in dysadherin-modified cells treated with shYAP, verteporfin, or YAP5SA. g ChIP-qPCR showing TEAD2 binding to the PD-L1 promoter region in dysadherin-OE PLC/PRF/5 cells. ChIP-Re-ChIP analysis showing co-occupancy of TEAD2 and YAP on the PD-L1 promoter in SK-Hep1 cells. Sequential immunoprecipitation was performed first with an anti-TEAD2 antibody, followed by an anti-YAP antibody. IgG served as a negative control. h Immunofluorescence analysis of PD-L1 and PD-1 binding in dysadherin-OE and KD cells using PD-1-Fc fusion protein staining. Scale bar = 100 μm. Flow cytometry analysis of CD69 expression (i) and the measurement of IFN-γ secretion (j) in Jurkat T cells co-cultured with dysadherin-modified HCC cells, treated with verteporfin or YAP5SA under CD3/CD28 stimulation. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests and one-way ANOVA with Dunnett’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001