Figure 3
Expression and splicing analysis of the pre-excised and post-excised lines. (A) Quantitative PCR analysis showed that integration into the PRPF39 gene caused a statistically significant increase in transcript expression, in which the increased expression was abrogated upon Adeno-Cre-PuroR-mediated hSTEMCCA excision. No statistical differences were seen between all control induced pluripotent stem cell (iPSC) lines, and all fibroblast lines displayed very low expression of PRPF39, indicating that this gene is associated with pluripotency. (B) To confirm that proper splicing of the transcript is taking place, primers against exons 4 + 5 show that no cryptic splice sites were being introduced and that, upon excision, the transcript correctly spliced itself. P <0.05. ESC, embryonic stem cell; Multiple Int., multiple integration; Syn., synthetic.