Figure 3.
Figure 3. KO of NOTCH4 by CRISPR-Cas9 promotes megakaryopoiesis. (A) Schematic illustration of the human NOTCH4 locus, showing the location of the gRNA binding site and the protospacer adjacent motif (PAM) sequence (red). (B) DNA sequences of the NOTCH4 WT and mutant clones. Red dots represent deleted bases, and the red lowercases are inserted sequences. The column on the right indicates the number of inserted or deleted bases. (C) Representative FACS analysis of the percentage of HSPCs (CD34+CD45+), megakaryocyte progenitors (MKPs) (CD34+CD41+), and MKs (CD41+CD42+) on day 14 in hematopoietic cells derived from NOTCH4−/−, NOTCH4+/−, or WT cells. (D) Bar graph depicting fold changes of the cell numbers of HSPCs, MKPs, and MKs on day 14 in hematopoietic cells derived from NOTCH4−/−, NOTCH4+/−, or WT cells. Data are shown as means ± SEM, n = 3. **P < .01; ***P < .001.

KO of NOTCH4 by CRISPR-Cas9 promotes megakaryopoiesis. (A) Schematic illustration of the human NOTCH4 locus, showing the location of the gRNA binding site and the protospacer adjacent motif (PAM) sequence (red). (B) DNA sequences of the NOTCH4 WT and mutant clones. Red dots represent deleted bases, and the red lowercases are inserted sequences. The column on the right indicates the number of inserted or deleted bases. (C) Representative FACS analysis of the percentage of HSPCs (CD34+CD45+), megakaryocyte progenitors (MKPs) (CD34+CD41+), and MKs (CD41+CD42+) on day 14 in hematopoietic cells derived from NOTCH4−/−, NOTCH4+/−, or WT cells. (D) Bar graph depicting fold changes of the cell numbers of HSPCs, MKPs, and MKs on day 14 in hematopoietic cells derived from NOTCH4−/−, NOTCH4+/−, or WT cells. Data are shown as means ± SEM, n = 3. **P < .01; ***P < .001.

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