Figure 4.

Enhanced interferon responses characterize infiltrating macrophages and monocytes in tumors from Lyz2-Cre/Myd88^fl/fl mice. scRNAseq was performed on CD45⁺ cells isolated from untreated tumors or 3d post-RT in Lyz2-Cre/Myd88^fl/fl mice and Myd88^fl/fl mice (n = 4 mice per group). (A) Volcano plot of differential gene expression in macrophages and monocytes (Itgam⁺Csf1r⁺Adgre1⁺ cells) that are upregulated in Lyz2-Cre/Myd88^fl/fl mice (right, orange) or upregulated in Myd88^fl/fl mice (left, blue). (B) Violin plots of select genes from (A). (C) Canonical pathway analysis using Ingenuity Pathway Analysis extrapolating likely pathways activated in Lyz2-Cre/Myd88^fl/fl mice treated with 16 Gy RT based off of significant differential gene expression from (A). (D) Upstream analysis based off of significant differential gene expression from (A) using Ingenuity Pathway Analysis, which identifies likely regulators that explain differential gene expression patterns observed in Lyz2-Cre/Myd88^fl/fl mice (top, orange) or Myd88^fl/fl mice (bottom, blue). (E) Quantitation of TNFα (left), IL-10 (center), and IL-6 (right) in supernatants of BMMΦs of the indicated genotype 24 h after treatment with 100 ng/mL LPS as measured by the ProcartaPlex platform. Significance of differential gene expression in (A) was defined as fold change > 1.5 and p < 0.1. Significance in (E) was assessed by an unpaired, two-sided t-test. * p < 0.05, ** p < 0.01, **** p < 0.0001.