Fig 3. The effect of HCQ and AZ on the presence of CD8⁺TCF1⁺ progenitor TILs.

Mice were implanted with B16 tumours as described in Fig 1. Panel A: viSNE profiles of total CD8⁺ TILs. The figure shows the presence of the upper cluster of TILs expressing moderate to high levels of CD8, PD-1 and CD44 (island (i)), a second island (ii) expressing moderate levels of TCF-1⁺ with no PD-1; and island (iii) that co-expresses PD-1⁺ and TCF-1⁺. Panel B: Percentage of CD8⁺ TCF-1⁺ PD-1^- TILs as a percent of total CD8+ TILs. Panel C: Ratio of CD8⁺ CD44⁺TCF-1⁺ PD-1^- TILs relative to tumor volume. Panel D: Ratio of CD8⁺ TCF-1⁺ PD-1⁺ TILs relative to tumor volume. Panel E: Ratio of CD8⁺ CD44⁺ PD-1⁺TCF-1⁺ progenitor TILs relative to tumor volume. A comparison of tumor volumes in mice where HCQ blocked the response to anti-PD-1 (big tumors) and mice where HCQ failed to block the response to anti-PD-1 (small tumors). Panel F: MFI of TCF-1 expression CD8⁺ CD44⁺TCF-1⁺ PD-1⁺ TILs in response to anti-PD-1 alone or in combination with HCQ or HCQ/AZ. Panel G: MFI of PD-1 expression CD8⁺ CD44⁺TCF-1⁺ PD-1⁺ TILs in response to anti-PD-1 alone or in combination with HCQ or HCQ/AZ. Panel H: MFI of T-bet expression CD8⁺ CD44⁺TCF-1⁺ PD-1⁺ TILs in response to anti-PD-1 alone or in combination with HCQ or HCQ/AZ. Panel I: MFI of Ki67 expression CD8⁺ CD44⁺TCF-1⁺ PD-1⁺ TILs in response to anti-PD-1 alone or in combination with HCQ or HCQ/AZ. Panel J: MFI of CD8 expression CD8⁺ CD44⁺TCF-1⁺ PD-1⁺ TILs in response to anti-PD-1 alone or in combination with HCQ or HCQ/AZ.