Supplemental Figure 6 from Increased Glucose Availability Sensitizes Pancreatic Cancer to Macrophage-Targeting Immunotherapies

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Supplemental Figure 6 - Flow cytometry gating strategy for immune profiling of pancreatic tumor–bearing mice. Representative flow cytometry plots illustrating the sequential gating strategy used for the identification of lymphoid and myeloid populations in pancreatic tumor samples (indicated as PAN_HG). Briefly, total events were first gated based on forward- and side-scatter (FSC-A vs SSC-A) to exclude debris, followed by singlet discrimination using FSC-A vs FSC-H. Viable leukocytes were identified and gated based on forward- and side-scatter properties, after which CD3+ T cells were selected and further subdivided into CD4+ and CD8+ T cell populations. Macrophages were gated on F4/80+ and were further characterized based on expression of iNOS and Arginase-1 to define inflammatory (iNOS+) and immunosuppressive (Arginase-1+) phenotypes. Additional lineage markers, including NK1.1, B220, CD140α, and PD-L1, were used to delineate specific immune subpopulations as indicated. Percentages shown in each gate represent the proportion of parent populations. All gates were set based on FMO controls and applied uniformly across all samples.

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