Figure 3
- ID
- ZDB-FIG-220522-15
- Publication
- Solman et al., 2022 - Inflammatory response in hematopoietic stem and progenitor cells triggered by activating SHP2 mutations evokes blood defects
- Other Figures
- All Figure Page
- Back to All Figure Page
(A) Schematic representation of the experimental procedure. At 5 days post fertilization (dpf), caudal hematopoietic tissues (CHTs) from Shp2wt, Shp2D61G/wt, and Shp2D61G/D61G embryos in Tg(cd41:GFP, kdrl:mCherry-CAAX) background were isolated. Cells were dissociated and separated by fluorescence-activated cell sorting (FACS), based on cd41:GFPlow expression, prior to single-cell RNA sequencing, as described in the Materials and methods section. (B) Number of cells of distinct genotypes used in single-cell RNA sequencing analysis. (C) Combined t-distributed stochastic neighbor embedding (t-SNE) map generated using the cells of all three genotypes (Shp2wt, Shp2D61G/wt, and Shp2D61G/D61G). Single cells from four major clusters are marked in dark green, blue, pink, and green, and their identities based on marker gene expression are indicated. Minor clusters are marked in gray. (D) Barplots showing the percentage of cells of Shp2wt, Shp2D61G/wt, and Shp2D61G/D61G genotype in distinct clusters. (E) Cells of distinct genotypes Shp2wt, Shp2D61G/wt, and Shp2D61G/D61G are visualized in t-distributed stochastic neighbor embedding (t-SNE) maps in red. (F) t-SNE maps showing log2-transformed read-counts of pu.1. (G) Representative images of the WISH staining for pu.1 expression in 5 dpf Shp2wt and Shp2D61G zebrafish embryos. Scale bar, 100 μm. (H) Expression of the pu.1 marker scored as low, mid, and high. (I) t-SNE maps showing log2-transformed read-counts of alas-2. (J) Representative images of the WISH staining for alas-2 expression in the tail region of 5 dpf Shp2wt and Shp2D61G zebrafish embryos. Scale bar, 100 μm. (K) Expression of the alas-2 marker scored as low, mid, and high. (H,K) Number on bars: number of embryos.
|