Figure S3.

General Dyes Enhance Hematopoietic Cell Type Segregation in FACS Space and Allow Purification of Eosinophils and HSPCs on BD FACSJazz, Related to Figure 3

(A) t-SNE map of zebrafish stained WKM TD2 generated on BD FACSJazz. Single cells are colored based on cell type.

(B) t-SNE map of zebrafish stained WKM TD3 generated on BD FACSInflux. Single cells are colored based on cell type.

(C) Contour plots of stained WKM cells showing normalized sorting gates for eosinophils for WKM 8 experiment (representative example for WKM 4, WKM 8 and WKM 9 eosinophil enrichments) on BD FACSJazz. Sorted cells passed through gate 1 and gate 2. Percentages of events within each gate are indicated.

(D) t-SNE map of zebrafish WKM TD2 where eosinophils inside and outside of GateID gates are colored in red and blue respectively.

(E) Projection of the sorted GateID eosinophils in WKM 8 (representative example for WKM 4, 8 and 9 eosinophil enrichment experiments) in FSC Height versus SSC Height.

(F and G) Barplots and t-SNE maps showing the outcome of eosinophil enrichments for (F) WKM 8 and (G) WKM 9 on BD FACSJazz.

(H) Left panel: FSC Height versus CD41-EGFP dot plot of live singlet WKM cells. The CD41^low gate is represented in red. Right panel: projection of the CD41^low sorted cells in FSC Height versus SSC Height.

(I) Barplot indicating cell type percentages for sorted CD41^low cells. Percentage in the barplot indicates HSPC percentage in the sorted library.

(J) t-SNE map showing sorted CD41^low cells. Non HSPCs are represented in gray and HSPCs in red.