Figure 2.
Characterization of TME-macrophage functions upon chemotherapy on Eμ-TCL1 mice.Eμ-TCL1-Cd19Cre-TP53wt (Eμ-TCL1) and Eμ-TCL1-Cd19Cre-TP53fl/fl (Eμ-TCL1) leukemic mice (Cd45+Cd19+Cd5+ cells on blood >30%) were IP injected 10 mg/kg cyclophosphamide (CTX) or PBS for 24 hours. (A) Schematic representation of the experimental design (upper panel). Splenocytes were isolated from the spleens of each treatment group, followed by MACS B-cell depletion (Cd19− cells) before scRNA-seq. Integrated Uniform Manifold Approximation and Projection (UMAP) dimension reduction plot of all treatment groups (n = 21 782 cells) (center panel). Cells are colored by the clusters determined by cell type. Percentage and total cell number of each treatment group contributing to the integrated UMAP (lower panel). (B) Pie chart representing the proportion of each cluster across all genotypes and treatment groups (left panel). Heat map showing the cell marker gene expression profiles of all macrophage clusters (MΦ-1, MΦ-2, MΦ-3, and MΦ-4) across all genotypes and treatment groups (right panel). (C) Diagram representing the gene ontology (GO) term analysis strategy across both genotype and treatment groups. (D) Pie chart showing the percentage of genotype and treatment group significantly regulated genes (left panel). GO process/function/component table showing the number of significant GO terms per genotype and treatment group (right panel). (E) GGnet plot of chemotaxsis/myeloid/phagocytosis GO process terms enriched in the macrophage clusters together with notation of the genotype and treatment group of significant genes of the GO terms (black, Eµ-TCL1 PBS; gray, Eµ-TCL1 CTX; light red, Eµ-TCL1Tp53fl/fl; dark red, Eµ-TCL1Tp53fl/fl CTX; dark green, >1 significant group). (F) Ball plot showing the percentage of positive cells and average expression across all macrophage cluster cells per genotype and treatment group for key GO process genes (left panel). Bar graph showing the fold change treatment normalized (CTX/PBS) per genotype per macrophage cluster (MΦ-1, MΦ-2, MΦ-3, and MΦ-4) for the same genes as in the left panel (right panel). Significant differences calculated either in Seurat (B/D), GO Gorilla (D/E), or GraphPad (F), respectively. In all instances, adjusted P values from the respective methods were used (see “Materials and methods”). IP, intraperitoneal; MACS, magnetic-activated cell sorting.

Characterization of TME-macrophage functions upon chemotherapy on Eμ-TCL1 mice.Eμ-TCL1-Cd19Cre-TP53wt (Eμ-TCL1) and Eμ-TCL1-Cd19Cre-TP53fl/fl (Eμ-TCL1) leukemic mice (Cd45+Cd19+Cd5+ cells on blood >30%) were IP injected 10 mg/kg cyclophosphamide (CTX) or PBS for 24 hours. (A) Schematic representation of the experimental design (upper panel). Splenocytes were isolated from the spleens of each treatment group, followed by MACS B-cell depletion (Cd19− cells) before scRNA-seq. Integrated Uniform Manifold Approximation and Projection (UMAP) dimension reduction plot of all treatment groups (n = 21 782 cells) (center panel). Cells are colored by the clusters determined by cell type. Percentage and total cell number of each treatment group contributing to the integrated UMAP (lower panel). (B) Pie chart representing the proportion of each cluster across all genotypes and treatment groups (left panel). Heat map showing the cell marker gene expression profiles of all macrophage clusters (MΦ-1, MΦ-2, MΦ-3, and MΦ-4) across all genotypes and treatment groups (right panel). (C) Diagram representing the gene ontology (GO) term analysis strategy across both genotype and treatment groups. (D) Pie chart showing the percentage of genotype and treatment group significantly regulated genes (left panel). GO process/function/component table showing the number of significant GO terms per genotype and treatment group (right panel). (E) GGnet plot of chemotaxsis/myeloid/phagocytosis GO process terms enriched in the macrophage clusters together with notation of the genotype and treatment group of significant genes of the GO terms (black, Eµ-TCL1 PBS; gray, Eµ-TCL1 CTX; light red, Eµ-TCL1Tp53fl/fl; dark red, Eµ-TCL1Tp53fl/fl CTX; dark green, >1 significant group). (F) Ball plot showing the percentage of positive cells and average expression across all macrophage cluster cells per genotype and treatment group for key GO process genes (left panel). Bar graph showing the fold change treatment normalized (CTX/PBS) per genotype per macrophage cluster (MΦ-1, MΦ-2, MΦ-3, and MΦ-4) for the same genes as in the left panel (right panel). Significant differences calculated either in Seurat (B/D), GO Gorilla (D/E), or GraphPad (F), respectively. In all instances, adjusted P values from the respective methods were used (see “Materials and methods”). IP, intraperitoneal; MACS, magnetic-activated cell sorting.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal