American Society of Hematology

Figure 1.

$Figure 1. BGLT3 encodes a 1.6-kb transcript that is dynamically cotranscribed with fetal γ-globin genes in erythroid cells in vitro and in vivo. Expression of BGLT3 and γ-globin determined by qRT-PCR in mouse E10.5 yolk sac (YS) and E12.5 fetal liver (FL) (A), CD34+ umbilical cord blood cells during expansion (days 6 and 8) and differentiation in culture (days 11, 12, and 13) (B), and K562 cells during treatment with hemin (C). (D) RNA FISH analysis of fetal hemoglobin–expressing erythroid cells from patients with the Corfu deletion; scalebar, 5 µm. (E) BGLT3 RNA as a fraction of total RNA in K562 nuclear and cytoplasmic fractions determined by qRT-PCR with primer pairs across BGLT3 (panel G shows locations). Intron and exon GAPDH primers (GI, GE) served as control. (F) 5′ and 3′ RACE of messenger RNA from K562 cells; lane 1, RNA; lane 2, control; n = 2 biological replicates. (G) Schematic diagram of Aγ-globin and BGLT3. Black lines show location of primers for panels E and H and Figures 3 and 5. (H) PCR products generated with (+) or without (−) complementary DNA or genomic DNA (dn) from differentiated (day 11) CD34+ cells. Primer sequences listed in supplemental Table 1. het, heterozygous carrier; hom, homozygous patient; RT+/−, with or without reverse transcriptase.$

BGLT3 encodes a 1.6-kb transcript that is dynamically cotranscribed with fetal γ-globin genes in erythroid cells in vitro and in vivo. Expression of BGLT3 and γ-globin determined by qRT-PCR in mouse E10.5 yolk sac (YS) and E12.5 fetal liver (FL) (A), CD34⁺ umbilical cord blood cells during expansion (days 6 and 8) and differentiation in culture (days 11, 12, and 13) (B), and K562 cells during treatment with hemin (C). (D) RNA FISH analysis of fetal hemoglobin–expressing erythroid cells from patients with the Corfu deletion; scalebar, 5 µm. (E) BGLT3 RNA as a fraction of total RNA in K562 nuclear and cytoplasmic fractions determined by qRT-PCR with primer pairs across BGLT3 (panel G shows locations). Intron and exon GAPDH primers (GI, GE) served as control. (F) 5′ and 3′ RACE of messenger RNA from K562 cells; lane 1, RNA; lane 2, control; n = 2 biological replicates. (G) Schematic diagram of ^Aγ-globin and BGLT3. Black lines show location of primers for panels E and H and Figures 3 and 5. (H) PCR products generated with (+) or without (−) complementary DNA or genomic DNA (dn) from differentiated (day 11) CD34⁺ cells. Primer sequences listed in supplemental Table 1. het, heterozygous carrier; hom, homozygous patient; RT^+/−, with or without reverse transcriptase.

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