Fig. S6.

The gating strategy used to determine TAM recruitment and phenotype in Gl261 tumors. Single-cell suspensions were stained for the following markers: 7AAD (cell viability), CD45, F4/80, CD11c, and MRC1. Cells were first gated according to side-scattered area (SSC-A) and forward-scattered area (FSC-A). Further gating according to SSC-A and side-scattered width (SSC-W) and FSC-A and forward-scattered width (FSC-W) was used to identify single cells. 7AAD was used to identify dead cells. Viable cells then were gated on CD45 to identify CD45⁺dsRed⁻ tumor-infiltrating immune cells. CD45⁺ cells were further gated on F4/80 to identify CD45⁺F4/80⁺ TAMs. Finally, CD11c and MRC1 were used to identify specific TAM phenotypes: M1-like TAMs (CD11c⁺MRC1⁻), M2-like TAMs (CD11c⁻MRC1⁺), M1–M2 intermediate TAMs (CD11c⁺MRC1⁺), and double-negative TAMs (CD11c⁻MRC1⁻).