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Fig. 1 | Cell Regeneration

Fig. 1

From: GATA2−/− human ESCs undergo attenuated endothelial to hematopoietic transition and thereafter granulocyte commitment

Fig. 1

Generation of GATA2 −/− human ESCs. a Schematic overview of gene targeting strategy. For GATA2 knockout, a PGK promoter-neomycin cassette replaces exon 3 of the GATA2 locus. BglII sites, TALEN sites, primers (P1, P2), and probes (probe 1, probe 2) used for genomic PCR and Southern blot are indicated. b Genomic PCR for GATA2 targeting. PCR product by indicated primers is 0.76 kb from WT H1 cells and 2.44 kb from H1-GATA2 −/−. c, d Southern blot analysis of GATA2 targeted H1 cells. The genomic DNAs were digested by BglII and detected by indicated probes. One single band detected by probe 1 indicates that there was no random integration of drug cassette. e FACS analysis of the expression of OCT4, SSEA-4, TRA-1-60, and TRA-1-81 in H1 or H1-GATA2 −/− cells. In these and other flow cytometry diagrams, the black line stands for the control and the red line for the experimental plot unless otherwise indicated. f Teratoma formation of H1 or H1-GATA2 −/− cells. g Paired Pearson correlation analysis of global gene expression between WT and GATA2 −/− H1 cells. R Pearson correlation coefficient, TPM transcripts per million. h Karyotype analysis of the H1-GATA2 −/− cell line

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