Fig. 1

A zebrafish blastomere culture system identifies the role of the chemical drug BF170 in hematopoietic cell induction. (A) (Top) Schematic of gene expression detection in cmyb-EGFP labeled cells in zebrafish primary blastomere cultured clusters from Tg (cmyb: EGFP) embryos. (Middle) Gene expression in disassociated GFP-positive cells from cultured blastomere embryonic cells or from the whole embryos by qRT-PCR. (Bottom) The expression of marker genes in GFP-positive cells and in GFP-negative cells from the cultured cmyb-EGFP clusters; the gene expression level in GFP-negative cells was normalized to 1. (B) Chemical structure of BF170. (C) BF170 increased expression of all the tested blood-related genes in zebrafish blastomere cultured clusters. (D) The cell morphology (top) and flow cytometry analysis (bottom) of apelin receptor-positive (APLNR+) cells in human ES (H1) cells treated with DMSO (as a control), BF170, flupirtine maleate (FLU) or quazinone (QUA) at day 3. Graph shows the percentage of APLNR+ cells in different groups. (E) The cell morphology (top) and flow cytometry analysis (bottom) of H1 cells at day 6 of hematopoietic differentiation. Graph shows the percentage of CD31+CD34+ cells. Each experiment was repeated three times. Data are mean±s.d. **P<0.01, ***P<0.001. Scale bars: 20 μm in D,E; 10 μm in A.