Enhanced gene delivery using a serotype chimeric modified recombinant Ad.5/3. (A) Expression of CAR and CD46 in pancreatic cancer cells as measured by flow cytometry. Cells were stained with anti-CAR, anti-CD46, and unstained or isotype-specific primary antibody. (B) Western blotting of desmoglein-2 (DSG-2) expression. (C) mRNA levels of CAR, CD46, and DSG-2 determined by real-time PCR. RQ, relative quotient. (D) Pancreatic cell lines were infected with Ad.5-luc and Ad.5/3-luc and level of luciferase (luc) expression was measured 48 hours postinfection using a luminometer (CPS, counts per second). (E) Pancreatic cancer cell lines BxPC-3 and MIA PaCa-2 were pretreated with anti-CAR (α-CAR), anti-CD46 (α-CD46) and recombinant DSG-2 for 2 hours followed by Ad.5/3-luc infection, and luciferase expression was quantified using a luminometer (CPS, counts per second). Level of significance: *P < 0.05; **P < 0.01; ***P < 0.001.