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Cross-stage analyses of csf1ra and csf1rb in zebrafish using two strategies A. Visualization of the expression of csf1ra and csf1rb in the constructed zebrafish cross-stage reference. Cells are color-coded based on the expression levels of csf1ra and csf1rb. “csf1ra” represents cells with normalized csf1ra expression levels greater than 0, and “csf1rb” represents cells with normalized csf1rb expression levels over 0. “Both” indicates cells where the normalized expression levels both csf1ra and csf1rb exceed 0. B. Dot plot illustrating the expression of csf1ra and csf1rb across different blood cell types in the zebrafish cross-stage reference. Dot size represents the percentage of cells expressing the gene, while color indicates the average expression level. C. Visualization of csf1ra expression in the constructed zebrafish cross-stage reference across three developmental stages (AGM, CHT, and KM). Cells are color-coded by scaled expression of csf1ra, with the red circle highlighting the specific expression of csf1ra in macrophages across three developmental stages. D. Violin plot depicting the expression levels of csf1rb in different blood cell types of zebrafish cross-stage reference across zebrafish developmental stages. Significant differences (****, P < 0.0001, t-test) were observed in HSPCs, macrophages, and neutrophils between the AGM and CHT stages, as well as between the CHT and KM stages. For HSPCs and neutrophils, the relative proportion of csf1rb-positive cells (i.e., the number of cells with normalized csf1rb expression levels over 0 vs. the total number of cells at each stage for a specific cell type) between three developmental stages is shown in the upper pie chart. E. Dot plots showing the percentage of cells expressing csf1rb (left) and the average expression level of csf1rb (right) across zebrafish AGM, CHT, and KM in HSPCs and neutrophils of zebrafish cross-stage reference. F.–H. Top: UMAP visualization of csf1ra expression in individual datasets from zebrafish AGM [24] (F), CHT [36] (G), and KM [37] (H). The red circles indicate the specific expression of csf1ra in macrophages across the three developmental stages. Bottom: violin plot showing the expression levels of csf1rb in HSPCs and neutrophils across zebrafish developmental stages in individually selected datasets (****, P < 0.0001, t-test). Meanwhile, the relative proportion of csf1rb-positive cells (i.e., the number of cells with normalized csf1rb expression levels over 0 vs. the total number of cells in each cell type at a specific stage) between HSPCs and neutrophils is shown in the upper pie chart. NK, natural killer.
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