Notch signaling regulates megakaryopoiesis at specific stages. (A) Schematic diagram of hematopoietic and megakaryocytic differentiation; specific cytokines were indicated for differentiation. The time of 2 different GSIs (10 μM) added for inhibition of the Notch signaling pathway is indicated by stars. n.s., not significant. (B) Representative FACS analysis of the percentage of HSPCs (CD34⁺CD45⁺), MKPs (CD34⁺CD41⁺), and MKs (CD41⁺CD42⁺) on day 14 in hematopoietic cells derived from BC1 iPSCs, with addition of 10 μM GSIs (RO4929097 or DAPT) or DMSO (control) from day 2. (C) Bar graph depicting fold changes of the cell numbers of HSPCs, MKPs, and MKs on day 14 in hematopoietic cells derived from BC1 iPSCs, with addition of 10 μM GSIs (RO4929097 or DAPT) or DMSO (control) from day 2. Data are shown as the means ± SEM, n = 3. *P < .05; **P < .01; ***P < .001. (D) Bar graph depicting fold changes of the cell numbers of HSPCs, MKPs, and MKs on day 14 in hematopoietic cells derived from BC1 iPSCs, with addition of 10 μM GSIs (RO4929097 or DAPT) or DMSO (control) from day 8. Data are shown as the means ± SEM, n = 3. *P < .05; **P < .01. (E) Bar graph depicting fold changes of the cell numbers of HSPCs, MKPs, and MKs on day 14 in hematopoietic cells derived from BC1 iPSCs, with addition of 10 μM GSIs (RO4929097 or DAPT) or DMSO (control) from day 11. Data are shown as the means ± SEM, n = 3. ***P < .001.