Fig 5. Chemotherapy induced systemic response and altered tumor immune landscape assessment using single-cell transcriptomics, secretory protein, and flow cytometry analyses.

(A) 32-plex cytokine analysis of plasma from mice subject to vehicle or docetaxel treatment (n = 5). (B) Expression (%) of Ki67 in D2.0Rs in the mammary tumor tissue of DTX compared to VEH-treated mice. (C–K) Flow cytometric analysis showing tumor immune infiltrates including (C) neutrophils, (D) monocytes, (E) MDSCs, (F) total macrophages, (G) M1 macrophages, (H) M2 macrophages, (I) CD4 T cells, (J) CD8 T cells, and (K) Tregs comparing DTX treatment with VEH control (n = 3, each). Independent t test measurement shows statistical significance between treatment groups. (L) UMAP showing immune cells in the tumor tissue of VEH and DTX datasets merged (n = 2, each). (M) Percentage of cells from VEH control and DTX-treated mice, per cluster for immune cells. (N) Dot plots of selected markers from merged samples (top) and grouped by treatment, VEH and DTX (bottom). Dot size indicates the proportion of cells in each cluster expressing a gene and color shading indicates the relative level of gene expression. Flow gating strategy for this figure can be found in S8 Fig and raw flow cytometry data is deposited on Flow repository (FR-FCM-Z6J8). scRNA-seq source data is available on GEO (accession # GSE231350). Source data and source code can be found in S1 Data and S1 Code, respectively. DTX, docetaxel; UMAP, Uniform Manifold Approximation and Projection.