Fig. 4: Functional role of GZMA and F2R expression in tumor suppression.

A Correlation of GZMA and F2R expression with the infiltrated immune cells in 372 LIHC from TCGA. B The T cell-mediated tumor cell killing assay performed in a 96-well plate and a 24-well plate. a: HepG2 and Huh7 cells, infected with EGFP-Lv or F2R-Lv, and co-cultured with the CD3⁺ T cells infected with GZMA-rAd or RFP-rAd, respectively. b: HepG2 and Huh7 cells, infected with EGFP-sh-Lv or F2R-sh-Lv, and co-cultured with the CD3⁺ T cells infected with GZMA-sh-rAd or RFP-sh-rAd, respectively. The cells were stained with crystal violet and quantified at 570 nm in a spectrometer. C The cell counting kit-8 assay. The above-stated cell was co-cultured in a 96-well plate. Cells were incubated with 100 μl of a medium supplemented with 10 μl of the CCK-8 solution for 1 h. The absorbance was recorded at 450 nm using a microplate reader. D Western blots indicating the expression of activated caspase 3, F2R, and GZMA in the above-stated co-cultured cells. E The Calcein-AM/PI double staining assay. The above-stated cells were co-cultured in a 24-well plate. The apoptotic cells stained positively with propidium iodide (red), while the living cells were stained positively with Calcein AM (green). The PI and Calcein AM-positive cells were visualized using fluorescence microscopy. Scale bars: 80 μm. *p < 0.05, *p < 0.01, ***p < 0.001, ****p < 0.0001.