Transcriptome profiling of PD-L1 antibody–treated macrophages showed inflammatory phenotype, increased survival and proliferation, and decreased apoptosis. Transcriptome profiling of PD-L1 antibody–treated macrophages showed inflammatory phenotype, increased survival and proliferation, and decreased apoptosis. Macrophages were treated with medium only, irrelevant isotype antibody, or PD-L1 antibody for 24 hours prior to RNA extraction for RNA sequencing analysis. The PCA plot (A) depicts the relationship and grouping of the samples based on global gene expression with medium only in red, isotype antibody in blue, and PD-L1 antibody in yellow. The volcano plot (B) shows the number of genes upregulated and downregulated on PD-L1 antibody–treated macrophages compared with isotype antibody–treated macrophages for P value with FDR ≤ 0.05 and fold change ≤ −2 (left, green) or ≥ 2 (right, red). Gene Ontology enrichment analysis was used to classify all significantly upregulated (C) and downregulated (D) genes into biological processes, with the enrichment score on the x-axis. IPA was next used to identify altered signaling pathways (E) with the yellow line depicting statistical significance, and upregulated values in red and downregulated values in green. F, The top 25 upregulated (left, red) and downregulated (right, green) genes. These data were generated using macrophages from 9 mice, with 3 mice in each treatment group. Genevieve P. Hartley et al. Cancer Immunol Res 2018;6:1260-1273 ©2018 by American Association for Cancer Research