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Fig. 5 Notch signaling mediates the effects of BF170 on hematopoietic stem and progenitor cell induction. (A) Notch signaling, nitric oxide (NO) signaling and ciliary parts were enriched for differentially expressed genes (DEGs) in flk1+ cells from BF170-treated zebrafish embryo. (B,C) BF170 induced increased NICD protein levels (B) and increased Notch reporter GFP signaling (C) in the whole embryos. Images on the right in C are magnified views of the images on the left. Graphs in B and C quantify levels of NICD protein (B) and GFP-positive signals (C) in control and BF170-treated embryos. (D) BF170-induced increased expression of dll4, notch1a and notch3, and of Notch targets hey1, hey2, her1, her2 and her5 in flk1+cmyb+ double-positive cells. (E) The Notch inhibitor DAPT blocked BF170-induced increases in cmyb expression in zebrafish embryos. (F) Loss of notch3 blocked the increased cmyb expression induced by BF170. (G) Loss of jag1a suppressed the increased cmyb expression induced by BF170. Red arrowheads indicate positive signals. (H-J) Quantification of cmyb expression in embryos from different groups in E-G, respectively. Images are lateral views with anterior to the left and dorsal upwards. In E-G, Nchanged/Ntotal in the bottom right corner of each image indicates embryos with changed expression/total tested embryos. n values indicate the number of embryos with changed expression in each group. Each experiment was repeated three times. Data are mean±s.d. In the box and whisker plots, the boxes indicate mean and the error bars indicate s.d. Whiskers indicate min to max values and all data points are shown. *P<0.05, **P<0.01, ***P<0.001. Scale bars: 100 μm in C (20 μm in higher magnifications); 100 μm in E-G.