Extended Data Fig. 6: Validation, single cell RNA sequencing, and reference mapping of Arg1ZsGr mice after MI. | Nature Cardiovascular Research

Extended Data Fig. 6: Validation, single cell RNA sequencing, and reference mapping of Arg1ZsGr mice after MI.

From: Hypoxia sensing in resident cardiac macrophages regulates monocyte fate specification following ischemic heart injury

Extended Data Fig. 6

a, Representative 20x confocal images of ARG1 IHC in the infarct and remote zone of Arg1ZsGr mice 2, 7, 30 days after I/R. Quantification of IHC is displayed as the percentage of ZsGr+ cells that are ARG1+ZsGr+. N = 4 vs 4 vs 3. Sensitivity of Arg1ZsGr mice 2 days after I/R within the infarct, quantified as the percentage of ARG1+ cells that are ZsGr+ or ZsGr−. N = 4 vs 4. b, Representative 20x confocal images of VSIG4 IHC in the infarct and remote zone of Arg1ZsGr mice 2, 7, 30 days after I/R. Quantification of IHC is displayed as the percentage of ZsGr+ cells that are VSIG4+ZsGr+. N = 3 vs 4 vs 3. c, FACS gating strategy for isolation of ZsGr− and ZsGr+ monocytes and macrophages from Arg1ZsGr mice 2 and 30 days after I/R. d, Violin plots representing scRNAseq data post quality control. Cells with greater than 500 and less than 5000 genes (nFeature_RNA), read counts less than 20,000 (nCount_RNA), and proportion of transcripts mapping to mitochondrial genes less than 10 percent (prompt) were used. e, Arg1 expression in the ZsGr− and ZsGr+ libraries at day 2 and 30. f, Stacked bar graph representing the proportion of each cell population in the data split between ZsGr− and ZsGr+ cells 2 and 30 days after I/R. g, Heat map of mapping scores for each cell population in the Query (Arg1ZsGr) plotted against the Reference MI data set. h, Z-score profiles from the reference data set (Extended Data Fig. 2f) compared to each subpopulation in the Arg1ZsGr data set represented as a dot-plot. i, Reference UMAP plots split between ZsGr− and ZsGr+ libraries 2 and 30 days after I/R after mapping to reference MI data set. The reference MI scRNAseq data set was the same as the one displayed in Extended Data Fig. 2. P-values are determined using the ordinary one-way ANOVA using Tukey’s multiple comparisons test. For N, each data point is an individual mouse. Data are presented as mean ± SEM.

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