Figure 4
Transcriptional profiling of Kidney-resident macrophages (KRM) demonstrates upregulation of Angiogenesis and Wound healing pathways in stenotic kidneys. (A) Ischemia-associated gene expression changes in RAS compared to Sham kidneys in CD11chiMϕ, CD11cloMϕ and KRM, respectively, displayed as volcano plots of individual genes, where fold-change between populations is plotted on the x-axis and significance on the y-axis. Genes upregulated >2-fold are in red, and genes downregulated >2-fold in blue. In RAS-CD11chiMϕ, 934 genes show changes in expression, with 766 up- and 168 down-regulated; while in RAS- CD11cloMϕ, 307 genes change, 241 up- and 66 down-regulated; and KRMs display the greatest changes in stenotic injury, with 3162 DEGs; 1506 are up- and 1656 down-regulated. (B) Enrichment analysis of biological process ontology in CD11chiMϕ, CD11cloMϕ and KRM. Top upregulated (top, red) and downregulated (bottom, blue) pathways in macrophage populations isolated from stenotic compared to Sham kidneys (pathway enrichment P < 0.05). (C) Contributions of different macrophage populations to injury response shown as log2 fold-change in expression of CD11chiMϕ, CD11cloMϕ and KRM. Each dot represents a gene that is up (red) or downregulated (blue) in RAS > 2-fold compared to sham control. (D) Gene signatures that are upregulated (angiogenesis, wound healing, and inflammation) or downregulated (interferon signature) in KRM in injury presented as heat-maps with hierarchical clustering. Mean values per Mϕ populations are shown. The z-score-based color-scale shows gene expression standard deviations below (blue) or above (red) the population mean. (E) Expression of genes involved in angiogenesis, and anti-inflammatory response presented as fold change over respective Sham and validated by RT-qPCR for individual macrophage samples from Sham and RAS mice. RAS KRM n = 3, RAS CD11chiMϕ, n = 3 and RAS CD11cloMϕ, n = 3; *P < 0.01 RAS-KRM vs RAS-CD11chiMϕ and *P < 0.01 RAS-KRM vs RAS-CD11cloMϕ. #P < 0.01 RAS vs respective Sham.
