Fig. 6: The molecular mechanism underlying the role of F2R in GZMA expression-based tumor suppression.

A Western blots indicating the expressions of activated caspase3, JAK2, p-JAK2, and F2R in the CD3⁺ cells co-cultured with the Huh7 cells infected with F2R-rAd, LDPRSFLL mutated motif F2R-rAd, and LDPRSFLL deleted motif F2R-rAd. B Western blots indicating the expressions of activated caspase3, JAK2, p-JAK2, and F2R in the CD3⁺ cells co-cultured with the Huh7 cells incubated with the LDPRSFLL motif-specific inhibitor (either SCH530348 or SCH79797). C The T cell-mediated tumor cell killing assay. The above-stated cells were stained with crystal violet and quantified at 570 nm in a spectrometer. D The Calcein-AM/PI double staining assay. The above-stated cells were co-cultured in 24-well plates. The apoptotic cells stained positively with propidium iodide (red), while the living cells stained positively with Calcein AM (green). The PI and Calcein AM-positive cells were visualized using fluorescence microscopy. Scale bars: 80 μm. E The nuclear extraction assay. The nuclear protein and the cytoplasmic protein in the above-stated cells were extracted using the nuclear extraction kit. Western blots indicated the level of STAT1 in the nucleus and the cytoplasm. F Fluorescence confocal assay results illustrating the subcellular localization of STAT1 (red) in the above-stated co-cultured cells. The nuclear DNA was stained with DAPI (blue). Scale bars: 15 μm.