Fig. 1

IRFA for HCC cells promotes autophagy with the emergence of an immunosuppressive microenvironment. A Western blot of LC3B and P62 after SMMC7721 and Huh7 cells heated at different temperatures (43, 45, 47, and 49 °C) for 15 min. B Immunoblotting of LC3B and P62 in SMMC7721 and Huh7 cells heated at 47 °C for different periods of time (0, 5, 15, 30, 45, and 60 min). C Representative CLSM images of SMMC7721 and Huh7 cells transfected with mRFP-eGFP–LC3 plasmid under heat stress (arrow pointing towards green spots, red spots, and yellow spots). Scale: 20 µm. D Schematic diagram of the establishment of IRFA subcutaneous HCC model. E Immunoblotting of LC3B and P62 in tumors before, 3 days, and 8 days after IRFA. F Representative CLSM images of LC3B, P62, CD4, and CD8 in tumors before, 3 days, and 8 days after IRFA. Scale: 100 µm. G, H Representative flow cytometry plots and quantitative assessments of CD4⁺ T cells, CD8⁺ T cells, CD11c⁺DCs and CD11b⁺/F4/80⁺ TAMs (of CD45⁺ gate) in tumors before, 3 days, and 8 days after IRFA (n = 3). I Quantification of TNF-α, IFN-γ, TGF-β, IL-6 in mice serum before, 3 days, and 8 days after IRFA by ELISA (n = 4). The data were expressed as mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. IRFA, Incomplete Radio Frequency Ablation; CLSM, Confocal laser scanning microscope; DCs, Dendritic cells; TAMs, Tumor-associated macrophages; ELISA, Enzyme-Linked Immunosorbent Assay