Fig. 1.
Fig. 1. Characterization of immortalized EKLF−/−cell lines. / (A) Appearance of transduced EKLF−/− colonies in methylcellulose after 5 days of culture in Epo (2 U/mL). Typical colony-forming unit–erythroid and a large erythroid colony are indicated by arrow and arrowhead, respectively (magnification × 40). (B-E) Cytocentrifuge specimens of the EKLF−/− cell line B1. (B) May-Grünwald-Giemsa stain showing erythroblast morphologic features. (C) Stain for hemoglobin usingo-dianisidine and Harris hematoxylin as counterstain. (D) Expression of human γ-globin by direct immunofluorescence with an FITC-conjugated antibody raised against hemoglobin F. (E) The same field as in panel D but photographed after UV excitation to demonstrate blue nuclear fluorescence (DAPI). (F,G) Cytocentrifuge specimens of a negative control cell line, J2E, which contains no human DNA sequences and shows no detectable γ-globin. The slide was treated as described for panels C and 1D.

Characterization of immortalized EKLF−/−cell lines.

(A) Appearance of transduced EKLF−/− colonies in methylcellulose after 5 days of culture in Epo (2 U/mL). Typical colony-forming unit–erythroid and a large erythroid colony are indicated by arrow and arrowhead, respectively (magnification × 40). (B-E) Cytocentrifuge specimens of the EKLF−/− cell line B1. (B) May-Grünwald-Giemsa stain showing erythroblast morphologic features. (C) Stain for hemoglobin usingo-dianisidine and Harris hematoxylin as counterstain. (D) Expression of human γ-globin by direct immunofluorescence with an FITC-conjugated antibody raised against hemoglobin F. (E) The same field as in panel D but photographed after UV excitation to demonstrate blue nuclear fluorescence (DAPI). (F,G) Cytocentrifuge specimens of a negative control cell line, J2E, which contains no human DNA sequences and shows no detectable γ-globin. The slide was treated as described for panels C and 1D.

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