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. Author manuscript; available in PMC: 2011 Feb 23.
Published in final edited form as: Biochemistry. 2010 Feb 23;49(7):1507. doi: 10.1021/bi9016846

FIGURE 5.

FIGURE 5

Long-term expression of DsRed in individual clones. (A) Flow cytometry analysis of DsRed expression in clones at 1 month (upper panel) and at 6 months (lower panel). The clone number is indicated above and the percent DsRed+ cells indicated in the upper left corner of each plot. (B) Analysis of DsRed expression by flow cytometry in two individual subclones 51-9 (65.7% DsRed+) and 51-11 (4.0% DsRed+), both of which were derived at 6 months from the same DsRed-positive cell clone 51 which had dropped from 74% of the cells expressing DsRed at 1 month to 5% at 6 months. Clones were stained for the erythroid specific marker CD235a, and the percentage of single or dual positive cells was determined by FACS analysis. The percentages of the population positive or negative for DsRed (vertical axis), and/or CD235a (horizontal axis) are indicated in the upper or lower corner of the relevant quadrant of the FACS plot. (C) Ligation-mediated PCR analysis of the genomic insertion sites in the parental clone 51 and the two subclones 51-9 and 51-11. Clone 51, sub-clones 51-9, and 51-11 gave the same product bands, a and b, which when sequenced were determined to be the same flanking host genomic sequences as those flanking the dual insertion sites in the parental clone 51 (Figure 2A). M, DNA ladder.