Figure 2.

Comparison of single-stranded (ss) and self-complementary (sc) adenovirus-associated virus (AAV) vectors in cell culture. HeLa S2 cells were transduced with AAV serotype 6 particles of the vectors AAV-EGFP 1xsU7 ss and scAAV 1xsU7 at a concentration of 500 vg/cell and analyzed for the splicing of the SMN2 minigene present in these cells by reverse transcriptase-polymerase chain reaction (RT-PCR) and agarose gel electrophoresis (a) and for the concentration of U7-ESE-B RNA by RT-quantitative PCR (RT-qPCR) (b). The RT-PCR for SMN2 splicing produces amplicons containing and lacking exon 7 as well as a more slowly migrating band corresponding to heterodimers (hd). All samples shown are from a single experiment with duplicate assays for each variable. The samples were processed together but analyzed on four different agarose gels. The RT-qPCR data have been compiled from three biological replicates analyzed with duplicate measurements. The curves were fitted with a third order polynomial function contained in the Excel program. The levels of 7SK snRNA were used for normalization.