Extended Data Fig. 9: Myeloid cell composition in B16-F10 tumors after IMQ therapy.
a, Flow cytometric gating strategy for lymphoid cells (see Fig. 7c, d) and myeloid cells (see Extended Data Fig. 9b–h). Cells were gated as follows: Within the viable cell gate: Immune cells (CD45+), T cells (CD3+, TCRβ+) CD4 or CD8α positive, B cells (-T cells, CD19+, B220+), Neutrophils (-B cells, Ly6-G+, CD11b+), Macrophages (-Neutrophils, CD64+, CD11b+, F4/80+), Monocytes (-Macrophages, Ly6-C+, CD11b+), pDCs (-Monocytes, BST-2+, B220+), DCs (-pDC, CD11c+, MHC-II+) XCR1 (Type I) or CD11b (Type II) positive and NK cells (-DC, NK1.1+). b, Stacked bar graph (horizontal) shows the frequency of myeloid cells in B16-F10 tumors after indicated treatment (Day 5). *P < 0.05. In b-e Control n = 5, IMQ: oral n = 4, topical n = 4, topical &oral n = 5; n is the number of mice from 1 experiment. c, Percentage of pDCs expressing PD-L1, CD80 or CD86, as assessed by flow cytometry in B16-F10 tumors treated as described in (b). d, Percentage of Type II DCs expressing PD-L1, CD80 or CD86, as assessed by flow cytometry at therapy end point (Day 5) as described in (b). e, Percentage of Type I DCs expressing PD-L1, CD80 or CD86, as assessed by flow cytometry, in B16-F10 tumors treated as described in (b). f, Frequency of CD8α T cells, positive for CD44 or PD-1 and TIM-3, was assessed by flow cytometry in B16-F10 at the ethical end point (Day 10). Mice were treated with IMQ orally or topically or both for 5 days. In f-h Control n = 5, IMQ: oral n = 3, topical n = 4, topical & oral n = 5; n is the number of mice from 1 experiment. g, Stacked bar graph (horizontal) shows the frequency of myeloid cells in B16-F10 tumors at the ethical end point (Day 10), as described in (f). h, Percentage of pDCs expressing PD-L1, CD80 or CD86, as assessed by flow cytometry in B16-F10 tumors treated as described in (f). Data are plotted as mean ± SEM. Dots in c-f and h represent biological replicates. P‐values were calculated using one-way ANOVA with Dunnett´s post-test (b-h).
