Integration of the human CFTR gene expression cassette into the AAVS1 locus. (A) Fluorescent images of IB3-1 cells transduced with 50 MOI or 100 MOI of HD-Ad-K18CFTR-TALEN vector; NC, untransduced cells. (B) Western blot for CFTR protein from cell lysate isolated 3 days post-transduction of 100 MOI of the HD-Ad-K18CFTR-TALEN vector. (C) Junction PCR analysis for IB3-1 cells transduced with the HD-Ad-K18CFTR-TALEN vector. Lane 1 shows the 4.6 kb product from the left-side junction and lane 2 shows the 4.7 kb product from the right-side junction. Lanes 3 and 4 are negative control PCR reactions for the left- and right-side junctions, respectively. (D) Restriction analysis of the junctional PCR products. Lane 1 and 2 are the left-side junction PCR product digested with EcoRV and AfeI, respectively; Lane 3 and 4 are the right-side junction PCR product digested with EcoRV and AfeI, respectively. (E) Restriction map for left-side and right-side junction PCR. (F) Right panel, Sanger sequencing around junction site of the PCR products.