Figure 2. Depletion of TAMs results in reduced ALDH^Bright TICs.

A) Analysis of leukocyte and ALDH^Bright TIC frequency in KCM tumors from mice treated for 21 days with vehicle, CSF1Ri1, CCR2i or in CCR2^−/− hosts. (i) The presence of CD11b⁺Ly6G⁻ Ly6C^LoF4/80^HiMHCII⁺ macrophage, CD11b⁺Ly6G^HiLy6C⁺ (G-MDSC/neutrophil), CD11b⁺Ly6G⁻ Ly6C^HiF4/80⁺MHCII⁺SSC^Lo monocyte (mono), CD11b⁺Ly6G⁻ Ly6C^LoCD11c^HiMHCII^Hi (M-DC), CD11b⁺Ly6G⁺Ly6C⁻ MHCII⁺ (basophil), and CD11b^LoLy6G⁻Ly6C⁻ CD11c^HiMHCII^Hi (Lymph-DC) subsets is depicted as the mean % of total live cells. (ii) ALDH^Bright TICs are depicted as the mean % of total live CD45⁻mCherry⁺ cells.

B) Analysis of macrophage subsets following CCR2 or CSF1R inhibition. (i) CD11b⁺CD3/19/49b⁻ Ly6G⁻Ly6C^LoF4/80⁺ macrophages were subdivided by MHCII and CD11c expression, and (ii) the mean frequency of each subset is displayed for all treatment groups. (iii) Relative expression of F4/80, CD206, and mCherry (indicator of phagocytosis) is depicted.

C) Flow cytometry analysis of TAMs and M-MDSCs infiltrating PAN02 tumors in mice treated for 4 or 8 days with vehicle, anti-CSF1, CCR2i, and/or CSF1Ri2 is depicted.

D) The mean frequency of macrophages and CD45⁻mCherry⁺ALDH^Bright TICs in Kras-INK tumors following 8 days of CSF1Ri treatment is depicted. Representative flow cytometry plots of mCherry⁺ ALDH^Bright tumor cells (Blue gate) are shown.

E) Quantitative-RT-PCR analysis in orthotopic Kras-INK tumor tissue following treatment with CSF1Ri for 14 days. Graph depicts the mean fold change compared vehicle.

Flow cytometry and quantitative-RT-PCR data depict the mean values from 5–10 mice ± SEM. * denotes statistically significant differences at p<0.05 (Mann-Whitney U-test).