Figure 2.

Expression of Aγ-globin gene in multicopy and single-copy ΔEnh/F lines. (A) Representative S1 nuclease protection analysis of expression of human Aγ versus mouse β^maj globin genes in adult blood RNA of single copy control lines, multicopy LCR-Aγ-ΔEnh/F-1, 2, and 3 lines, and the derived single copy (sc) lines analyzed. The type of protected fragments are indicated to the right of the panel. Controls used in this experiment included a 16.5 dpc embryo from HPFH2C line and an adult HPFH2C RNA sample (ad), expressing high levels of the Aγ-gene, as described by Katsantoni et al. (20). Expression levels were calculated by ImageQuant after correcting for probe-specific activities according to the ratio of 1:2.4 for Aγ:β^maj probes. (B) Quantitation of Aγ-globin expression levels of the multicopy LCR-Aγ-ΔEnh/F-1, 2, and 3 lines, and the derived single copy (sc) lines as a percentage of total β^maj globin gene expression. Data shown represent mean ± standard deviation from a total of three samples analyzed per each line. The values used in the chart are from Table 1.