Early CD206⁺ TAM depletion leads to a coordinated and indirect loss of NK, cDC1, and CD8 T cells in tumors. (A) Schematic representation of the experimental setup for early and late CD206⁺ TAM depletion in B78chOVA tumors using Mrc1(CD206)^{LSL-Venus-DTR} (WT) and Csf1r^Cre; CD206^{LSL-Venus-DTR} (DTR) mice. (B–H and J–P) Relative abundance of different immune populations in B78chOVA tumors as a percentage of CD45⁺ cells with (B–H) late and (J–P) early depletion regimens. (I and Q) Abundance of TAMs gated by CD206 expression, showing percentage depletion of CD206⁺ TAMs in (I) late and (Q) early depletion setting. (R–U) Representative flow cytometry plots showing CD206 versus MHCII expression in different intratumoral myeloid subsets in WT (red) and DTR (blue) mice in the (R) late and (S) early depletion regimens; relative abundance of (T) neutrophils, (U) cDC1, NK cells, and OT-I T cells in the early DTx administration setting with additional anti-Ly6G or isotype control treatment. (V–X) Relative abundance of (V) CD206⁺ cDC2, (W) cDC1, and (X) CD44⁺ CD8 T cells in the early DTx administration setting in WT and DTR^L (Lyz2(LysM)^Cre; CD206^{LSL-Venus-DTR}) mice. Bar graphs show mean ± SEM; data are representative of at least two independent experiments, each with at least three biological replicates per group; ***P < 0.001, **P < 0.01, *P < 0.05, ns = no significance by unpaired Student’s t tests or Mann–Whitney U test (B–Q and V–X) and Kruskal–Wallis test with post-hoc test correcting for false discovery rate (T and U).