Effect of hSET1A depletion on ESC mesoderm differentiation and hematopoiesis.
(A) Western blotting analysis of hSET1A protein levels in ESCs harboring scramble control or hSET1A specific shRNAs. (B) Hematopoietic differentiation assay. The scramble control or hSET1A KD ESCs were cultured in suspension without LIF to induce EB formation for 4 days and then cultured in the presence of SCF to induce hematopoietic differentiation for another 4 days. Shown are EBs after 8 days in culture. Scale bar, 100 µm. (C) to (E) Time course qRT-PCR analyses of the levels of mesoderm (C), endoderm (D), and ectoderm (E) markers in the scramble control and two hSET1A KD clones upon withdrawal of LIF. (F) FACS analysis of c-Kit and Tie2 expressing hemogenic endothelium population in the scramble control and hSET1A KD ESCs upon hematopoietic differentiation at day 6. (G) FACS analysis of CD41 and c-Kit expressing early hematopoietic stem and progenitor population in the scramble control and hSET1A KD ESCs upon hematopoietic differentiation at day 10.