Fig. 4 ∣. Loss of Elf5 alters the immune landscape in C3-T⁺ tumours.

a, FoxP3 staining shows an increased number of T_reg cells in C3-T⁺ Elf5^+/− tumours compared with C3-T⁺ Elf5^+/+ tumours. b, F4/80 staining shows an increased number of macrophages in C3-T⁺ Elf5^+/− tumours compared with C3-T⁺ Elf5^+/+ tumours. c, S100A8 analysis shows an increased number of myeloid cells in tumours from C3-T⁺ Elf5^+/− mice, d, CD8 staining shows decreased numbers of cytotoxic T cells in tumours from C3-T⁺ Elf5^+/− mice (arrowheads). In a-d, images of the stains are shown to the left, with quantification of the results to the right, and n = 3 individual tumours were used with a minimum of ten random fields per genotype for IHC quantification. The boxplot data represent medians, interquartile ranges and spikes to upper and lower adjacent values. Scale bars: 40 μm. e-h, Flow cytometry analyses (FACS) showing increased T_reg cells (CD45⁺CD3⁺CD4⁺FoxP3⁺; e) with no major change in F4/80⁺ (CD45⁺G4/80⁺; f), increased Gr1⁺ myeloid cells (CD45⁺CD11b⁺Gr1⁺; g) and decreased cytotoxic T cells (CD45⁺CD3⁺CD8⁺; h) in C3-T⁺ Elf5^+/− tumours compared with C3-T⁺ Elf5^+/+ tumours. All of the gates were drawn according to a published protocol⁵⁹. The scatter plots show the number per group of samples from flow analysis and the data are presented as medians ± s.e.m. Statistical significance was determined by two-tailed Mann–Whitney U-test (a-d and f-h) and two-tailed Student’s t-test (e).