Supplemental Figure 8 from Increased Glucose Availability Sensitizes Pancreatic Cancer to Macrophage-Targeting Immunotherapies

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Supplemental Figure 8 - Flow cytometric analysis of additional immune and stromal cell infiltration following single and combination treatments in pancreatic tumors. A.) Representative flow cytometry plots illustrating the effects of vehicle control, D30, PLX3397, PLX3397 + D30, PF-4136309, and PF-4136309 + D30 on stromal fibroblast populations within pancreatic tumors. Tumor-derived single-cell suspensions were first gated on viable cells based on forward- and side-scatter properties. The CD45− fraction was subsequently analyzed to identify CD140α+ fibroblasts, as shown by the indicated gates. Enlarged panels display representative SSC-A versus CD140α plots for each treatment group, with percentages of CD140α+ cells indicated. Bar graphs summarize the proportion of CD45+ immune cells and CD140α+ fibroblasts across treatment groups. B-C.) Representative flow cytometry plots and quantitative analyses showing the impact of vehicle control, D30, PLX3397, PLX3397 + D30, PF-4136309, and PF-4136309 + D30 on intratumoral B cells (B.) and NK1.1+ cells (C.). Representative contour plots display the frequencies of B220+ and NK1.1+ populations within the parent gates, with percentages shown in each panel. Bar graphs summarize the proportion of B220+ B cells and NK1.1+ cells across treatment groups. Data are presented as mean ± S.D. from independent biological replicates, with each dot representing an individual tumor. Statistical significance was assessed using one-way ANOVA with multiple-comparison correction. ns, not significant; * indicated p<0.05; ** indicated p<0.01; *** indicated p<0.001. All gates were defined using FMO controls and applied uniformly across all samples.

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