Fig. 3.

Validation of cellular hemoglobin expression. a Cells were cloned from single E-colonies, formed from HSPCs post CRISPR genome editing, and amplified after culturing for 10 days; left: pellet of amplified cells had reddish color; right: cell amplification. b Immunophenotyping analysis of cloned erythroid precursor cells by flow cytometry. c HPLC assay revealed a new peak for HbA* protein in the genome-edited erythroid progenitor cells (right panel); in contrast, a peak for HbS only was present in cloned E-colony cells that did not undergo genome editing as the experiment internal control (left panel). d Hemoglobin in the peripheral blood from SCD patient, SCT carrier, and normal person were used as HPLC assay standard controls. e HPLC retention times corresponding to peaks for HbA*/HbA and HbS. Notably, peaks for HbA* and natural HbA were detected at the same retention time because they have identical amino acid sequences