Fig. 3

Frequency and effector function of T-cells following selumetinib, anti-CTLA-4 and combination treatment in vivo. a Schema showing treatment schedule. b Immunohistochemical analysis of tumors for p-ERK 1 h following the last dose with 25 mg/kg selumetinib bid (3 doses in total, over 24 h) or vehicle control. Following 8 days of anti-CTLA-4, selumetinib or combination treatment, cells isolated from spleens and tumors were analysed by flow cytometry analysis. Spleens from non-tumor bearing BALB/c mice were also included in the analysis. c CD8⁺ T-cell and (d) CD4⁺ T-cell populations are presented as percentages of CD45⁺ cells. e Frequency of Foxp3⁺ CD4⁺ regulatory T-cells (Tregs) of total CD4⁺ T-cells. f Ratio of CD8⁺ T-cells to Tregs. Effects of treatment on the frequency of Ki67-positive cells (g) of total CD8⁺ T-cells, (h) CD4⁺ T-cells or (i) Tregs. Data points in scatter plots represent individual animals, treatment groups each contained 8 mice. Plotted are means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by unpaired t-test between indicated groups