Fig. 6: Combination treatment increases the amount of IFNγ⁺ CD8 T cells and their mobility in the TME.

a Flow cytometry quantification of IFNγ⁺ CD8 T cells (ctrl n = 10, aPD-L1 n = 10, aTGFb n = 5, combo n = 10). b Representative images of EMT6-mAPPLE tumors at 14 days after initiation of treatment (n = 8 per group). Blue, tumor cells; Red, IFNγ-YFP⁺ cells; Yellow, blood vessels; white dashed circles highlight IFNγ-YFP cells. c Density of IFNγ-YFP⁺ cells at three time points after initiation of treatment (n = 8 Ctrl, n = 7 combo (non-responders excluded); P < 0.0001 two-way ANOVA, mixed-effect analysis; mean and SEM are shown). d Correlation between tumor volume (y axis) and IFNγ-YFP⁺ cells density (x axis) (n = 8 per group). e IFNγ-YFP⁺ cell speed at day 7–9 after initiation of treatment (Control: n = 179 cells from 8 mice; Combination: n = 2310 cells from 7 mice (non-responders excluded); P < 0.0001, two-tailed Kolmogorov–Smirnov test). f IFNγ-YFP⁺ cells track length at day 7–9 after initiation of treatment (Control: n = 179 cells from 8 mice; Combination: n = 2,310 cells from seven mice (non-responders excluded); P value < 0.0001, two-tailed Kolmogorov–Smirnov test). g Tumor volume (y axis) of EMT6 tumors treated with anti-PD-L1, combo or combo plus IFNγ neutralizing antibody (a-IFNg) over time (x axis). Grey shade indicates treatment duration. Individual animal curves (grey lines) and group fit curves (thick solid lines) of the control group (black) and treatment groups (colored) are provided. CR complete responder, PR partial responder. Representative experiment of three (n = 10 for all groups). h Tumor volume (y axis) of EMT6 IFNGR1 KO tumors treated with anti-PD-L1 or combo over time (x axis). Grey shade indicates treatment duration. Individual animal curves (grey lines) and group fit curves (thick solid lines) of the control group (black) and treatment groups (colored) are provided. CR complete responder, PR partial responder (n = 10 for all groups). Source data are provided as a Source Data file. Ctrl = control (anti-GP120), aPD-L1 = anti-PD-L1, aTGFb = anti-TGFβ, combo = combination anti-PD-L1⁺ anti-TGFβ; Adj P values *<0.1, **<0.05 Dunn’s test (two-sided) with Benjamini–Hochberg multiple testing correction unless differently specified. Adjusted P values for (a): 0.0382 (ctrl vs combo), 0.0851 (aTGFb vs combo). a Whiskers represent the minimum and maximum (unless points extend 1.5 * IQR from the hinge, then shown as individual points), the box represents the interquartile range, and the center line represents the median.