Fig. 3: Knockout of UGCG and its effects on T cell anti-tumour in vitro and in vivo.

a Western blot analysis of the expression of UGCG in MC38-OVA with and without UGCG knockout. GAPDH was used as a protein loading control. The analysis was done thrice with biologically independent samples. b Cell proliferation of sgCONT or sgUGCG-MC38-OVA tumour cells by FACS analysis (n = 3). c Colony formation ability of MC38-OVA tumour cells with sgCONT or sgUGCG tumour cells on day 14 (n = 3). d Titration curves of MHC-I-specific antibodies on MC38-OVA cells. Flow cytometry charts of antibody stain as indicated by the arrow. MFI, mean fluorescence intensity (n = 3). e CD8⁺ T-cell killing assay of sgUGCG-MC38-OVA tumour cells cocultured in different ratios with CD8⁺ T cells isolated from OT-I mice. The number of viable sgUGCG-MC38-OVA tumour cells at the end point of the assay was determined and reported (n = 3). f FACS analysis IFN-γ, TNF-α, expression of CD8⁺ T cells after coculture with sgUGCG-MC38-OVA tumour cells at an effector-to-target ratio of 1:1 (n = 3). g, sgCONT- and sgUGCG-MC38-OVA cells grown in C57BL/6 mice(n = 10). About 2 × 10⁵ sgCONT- and sgUGCG-MC38-OVA cells were inoculated subcutaneously into syngeneic mice and monitored for tumour formation. h Phenotype and function of TILs from sgCONT- and sgUGCG-MC38-OVA tumours were detected by flow cytometry analysis at day 21 after tumour inoculation (n = 3). Surface levels of CD8⁺ T cells, IFN-γ, TNF-α and Tetramer on tumour-infiltrating CD8⁺ T cells of different groups were analysed. Bar colours (f, h): control sgRNA (red), sgRNA targeting UGCG (dark green). The data represent three independent experiments with similar results (b–f). Data are shown as the mean ± SEM. ns, not significant. P values in (b, c, f and h) were calculated by unpaired two-sided t test. P values in (e) were calculated by unpaired two-sided t tests. P values in (g) were determined by two-way repeated measures ANOVA. Source data and exact p values are provided as a Source Data file.