ZFIN Image: Solman et al., 2022, Figure 3

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Figure 3

Single-cell RNA sequencing of hematopoietic stem and progenitor cells (HSPCs) reveals myeloid bias in Shp2^D61G embryos.

(A) Schematic representation of the experimental procedure. At 5 days post fertilization (dpf), caudal hematopoietic tissues (CHTs) from Shp2^wt, Shp2^D61G/wt, and Shp2^D61G/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:GFP^low 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 (Shp2^wt, Shp2^D61G/wt, and Shp2^D61G/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 Shp2^wt, Shp2^D61G/wt, and Shp2^D61G/D61G genotype in distinct clusters. (E) Cells of distinct genotypes Shp2^wt, Shp2^D61G/wt, and Shp2^D61G/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 Shp2^wt and Shp2^D61G 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 Shp2^wt and Shp2^D61G 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.

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Figure 3

Figure 3

Single-cell RNA sequencing of hematopoietic stem and progenitor cells (HSPCs) reveals myeloid bias in Shp2D61G embryos.

Single-cell RNA sequencing of hematopoietic stem and progenitor cells (HSPCs) reveals myeloid bias in Shp2^D61G embryos.