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. Author manuscript; available in PMC: 2012 May 21.
Published in final edited form as: Nat Biotechnol. 2010 Dec 12;29(1):73–78. doi: 10.1038/nbt.1717

Figure 3.

Figure 3

β-globin expression in the erythroid progeny of single-vector-copy thal-iPS cell clones. (a) Expression of erythroid cell markers CD71 and glycophorin A (GPA) in the erythroid progeny of thal-iPS cell line 1.52. (b) β-globin expression in the erythroid progeny of 13 single-vector-copy thal-iPS cell clones assessed by qRT-PCR. Expression levels are expressed per gene copy, relative to the average endogenous β-globin expressed in the in vitro differentiated erythroid progeny of peripheral blood CD34+ cells from four healthy individuals and normalized to endogenous α-globin expression. hES: erythroid progeny of hES cell line H1, wt iPS: erythroid progeny of iPS cell line FDCT, derived from fibroblasts of an 11-year-old healthy individual30. Numbers below graphs depict thal-iPS clone numbers derived from lines thal1.52, thal2.1, thal5.10 and thal5.11. n: number of independent differentiations for each clone. UT: untransduced. Error bars denote s.e.m. (c) β-globin expression in the erythroid progeny of a subset of single-vector-copy thal-iPS cell clones and controls assessed by quantitative primer extension. βE: endogenous β-globin (80 bp); αE: endogenous α-globin (60 bp); βV: vector-encoded β-globin (84 bp). PB CD34+: erythroid cell derivatives of in vitro differentiated peripheral blood (PB) CD34+ cells from a normal donor; H1 hES: erythroid cell derivatives of the H1 hES line; wt iPS: erythroid cell derivatives of iPS cell line FDCT, derived from fibroblasts of a healthy individual; thal1.52 UT, thal2.1 UT: erythroid cell derivatives of untransduced lines thal1.52 and thal2.1, respectively; thal1.52-17, thal2.1-67, thal1.52-10, thal1.52-16, thal1.52-38, thal2.1-49, thal2.1-48, thal2.1-55, thal5.11-32, thal5.10-2: erythroid cell derivatives of the respective single-vector-copy thal-iPS clones. (d) Chromatograms of HPLC analysis of α- and β-globin expression in the erythroid progeny of clones thal5.10-2 and thal5.11-28. Cord blood, H1 hES and untransduced (UT) thal1.52 cells were used as controls. For quantification of these data, see Supplementary Table 7.