Extended Data Fig. 7: IL-4 deficiency impairs 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 Il4^−/−Tgfbr2^fl/fl PyMT (Il4^−/−; n = 4) and Il4^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Il4^−/− KO; n = 6) 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 Il4^−/−Tgfbr2^fl/fl PyMT (Il4^−/−; n = 5) and Il4^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Il4^−/− KO; n = 7) 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 Il4^−/−Tgfbr2^fl/fl PyMT (Il4^−/−) and Il4^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Il4^−/− KO) mice. Extravascular (EV) Fg deposition events (magenta arrows) were calculated from 1 mm² regions (n = 9 for Il4^−/−Tgfbr2^fl/fl PyMT and Il4^−/−Thpok-cre;Tgfbr2^fl/fl PyMT tumour tissues). Isolated CD31⁺ staining (yellow arrows) was counted from 1 mm² regions (n = 9 for Il4^−/−Tgfbr2^fl/fl PyMT and Il4^−/−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 Il4^−/−Tgfbr2^fl/fl PyMT (Il4^−/−) and Il4^−/−Thpok-cre;Tgfbr2^fl/fl PyMT (Il4^−/− KO) mice. The percentage of HPP⁺E-Cadherin⁺ regions over E-Cadherin⁺ epithelial regions was calculated from 1 mm² regions (n = 9 for Il4^−/−Tgfbr2^fl/fl PyMT and Il4^−/−Thpok-cre;Tgfbr2^fl/fl PyMT tumour tissues). Data are represented as the mean ± s.e.m. (biologically independent mice in a–d). Two-tailed unpaired t-test (a–d).

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