Fig. 7: ZEB1 in macrophages promotes abundance of CD8+ cells and concomitant tumor cell killing.

a Representative images and quantification of IHC for CD8 positive (+) (n > 57 fields of view (FOVs) of 7 mice) and cleaved Caspase 3+ cells (cl. Casp. 3 IHC scores from n > 7 mice) in s.c. CMT-93 (50-58 days-post injection (dpi), see Fig. 3a, b, c) and MC-38 tumors (13-19 dpi, also see Fig. 3d) in LysM^Ctrl and LysM^ΔZeb1 mice. Insets show higher magnification. b Percentage of s.c. CMT-93 tumor-bearing LysM^Ctrl, LysM^ΔZeb1 and NSG mice. Number of mice are indicated (tumor-bearing/ total). c Quantification of IHCs in MC-38 lung colonies in LysM^Ctrl and LysM^ΔZeb1 mice for CD8+ cells (n > 33 tumorous FOVs images of n > 6 mice) and cl. Casp. 3+ cells (IHC scores from n > 6 mice). d IHCs in KPC lung colonies in LysM^Ctrl and LysM^ΔZeb1 mice with representative images and quantification over time of CD8+ cells (n > 20 tumorous FOVs per condition; means +SEM; image at 10 dpi), of cl. Casp3+ cells (n > 10 tumorous FOVs per condition, means +SEM images at 14 dpi) and of CCL2 (IHC scores of colonies from n > 7 mice per condition; means ± SD; images at 10 dpi) e. Ki67 IHC in indicated lung colonies in LysM^Ctrl and LysM^ΔZeb1 mice with representative image of KPC colonies at 21dpi and quantification (IHC score of colonies from n > 6 mice per condition). f–h Precision cut lung slice cultures and therapeutic CD8 + T cell addition (TCA) of splenic CD8 + T cells with experimental setup created with Biorender (f), monitoring of KPC-mCherry total fluorescence (per lung area) as surrogate for cell growth on LysM^Ctrl and LysM^ΔZeb1 lung slices over time in the presence / absence of ACT (g) as well as the combination of CCR2 and CCR4 antagonists or DMSO as vehicle controls relative to ACT start (h). Means ± SD. ****p < 0.0001; ***p < 0.001 *p < 0.05; ns: not significant; Mann-Whitney (a, c; for CD8); Qui-square test (b); two-tailed t-test (a, c; for cl. Casp. 3; e); 2-way ANOVA (d, g, h).