Fig. 4: B3GNT2 disrupts ligand–receptor interactions between tumor and T cells.

a Cell survival against T cell cytotoxicity (top) and T cell IFNγ secretion (bottom) in A375 cells overexpressing B3GNT2 or GFP that have been treated with different concentrations of kifunensine. Kifunensine was used to pretreat A375 cells and was present during co-culture with T cells at E:T ratio of 3. Kifunensine-treated cells that were co-cultured with ESO T cells were compared to kifunensine-treated cells cultured in parallel without T cells to determine percent survival. N = 6. Two-tailed t tests were performed. b Tomato lectin IP of A375 cells overexpressing GFP or B3GNT2 followed by western blot for different B3GNT2 target proteins. 2% of the input and no lectin IP controls are shown. Data are representative of two independent experiments. c Western blots of A375 cells overexpressing B3GNT2 or GFP that were treated with kifunensine (KIF) or benzyl-2-acetamido-2-deoxy-α-D-galactopyranoside (BAG) to remove N- or O-glycosylation respectively. Data are representative of two independent experiments. d Histograms (top) and corresponding median fluorescence intensity (MFI; bottom) showing binding of recombinant T cell proteins to A375 cells measured by flow cytometry. A375 cells were overexpressing GFP or B3GNT2 and treated with KIF or BAG. N = 3. Two-tailed t tests were performed. e Schematic showing the tumor cell surface ligands and receptors modified by B3GNT2 to disrupt interactions with T cells that mediate cytotoxicity. All values are mean ± s.e.m. ns not significant. Source data are provided in Source Data 4.