Extended Data Fig. 6: IFN-γ deficiency does not impair cancer immunity triggered by blockage of TGF-β signalling in CD4⁺ T cells.

a, Representative flow cytometry plots of CD62L and CD44 expression and statistical analyses of the gated populations among CD4⁺FOXP3⁻ T cells (top panel) and CD4⁺FOXP3⁺ T cells (bottom panel) from Ifng^−/−Tgfbr2^fl/fl PyMT (Ifng^−/−; n = 3) and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Ifng^−/− KO; n = 4) mice. b, Representative flow cytometry plots of TCRβ, NK1.1, CD4, CD8 and FOXP3 expression and statistical analyses of the gated populations in tumour-infiltrating leukocytes from 23-week-old Ifng^−/−Tgfbr2^fl/fl PyMT (Ifng^−/−; n = 4) and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Ifng^−/− KO; n = 5) mice. c, Representative immunofluorescence images of fibrinogen (Fg, white), CD31 (red), cleaved Caspase 3 (CC3, cyan) and E-Cadherin (green) in comparable individual tumours with sizes around 8x8 mm in length and width from 23-week-old Ifng^−/−Tgfbr2^fl/fl PyMT (Ifng^−/−) and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Ifng^−/− KO) mice. Extravascular (EV) Fg deposition events (magenta arrows) were calculated from 1 mm² regions (n = 9 for Ifng^−/−Tgfbr2^fl/fl PyMT and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT tumour tissues). Isolated CD31⁺ staining (yellow arrows) was counted from 1 mm² regions (n = 9 for Ifng^−/−Tgfbr2^fl/fl PyMT and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT tumour tissues). d, Representative immunofluorescence images of a hypoxia probe (HPP, white), CD31 (red), cleaved Caspase 3 (CC3, cyan) and E-Cadherin (green) in tumours from Ifng^−/−Tgfbr2^fl/fl PyMT (Ifng^−/−) and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Ifng^−/− KO) mice. The percentage of HPP⁺E-Cadherin⁺ regions over E-Cadherin⁺ epithelial regions was calculated from 1 mm² regions (n = 9 for Ifng^−/−Tgfbr2^fl/fl PyMT and Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT tumour tissues). The shortest distance of HPP⁺ regions (magenta dashed lines) or CC3⁺ regions (yellow dashed lines) to CD31⁺ endothelial cells was measured in tumour tissues from Ifng^−/−Thpok-cre;Tgfbr2^fl/fl PyMT mice (n = 9). Data are represented as the mean ± s.e.m. (biologically independent mice in a–d). Two-tailed unpaired (a–d) or paired (d) t-test.

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