Fig. 4

Impaired hematopoietic differentiation of PRDM8^−/− iPSCs. a Scheme of two main transcripts of PRDM8 (NM_020226.3 and NM_001099403.2) and sites of genomic editing. Two pairs of guide RNAs (gRNAs) were designed targeting the intron/exon boundary at the start codon of both transcripts. b Genome editing was confirmed by gene expression analysis after 14 days of embryoid body assay (normalized to GAPDH and PRDM8 expression in the undifferentiated control cells). c Phase contrast pictures of EBs after 16 days of hematopoietic differentiation. The control EB produces hematopoietic progenitor cells, whereas this is not the case for the PRDM8^+/− clone and the PRDM8^−/− clone, which consistently revealed enhanced growth. Scale bar, 500 μM. d Knockout of PRDM8 resulted in a significantly lower number of hematopoietic progenitor cells. t test: * P < 0.05; ns, not significant. e Cytospins supported impaired hematopoiesis after PRDM8 knockout. Scale bar, 500 μM. f The colony forming unit (CFU) potential is lost in PRDM8 knockout clones. g Flow cytometry substantiates hematopoietic differentiation of the control iPSCs (read line: autofluorescence; blue line: with antibodies)