FIG. 8.

CAR gene expression is downregulated days after infection of BJAB cells. (A) Cellular RNA was extracted from Ad5dl309-infected BJAB cells at days 3 and 180 postinfection or from uninfected BJAB, A549, or KE37 cells transduced with the CAR gene (KE37-CARhi). Reverse transcription followed by PCR was used to detect mRNA for CAR and β₂-microglobulin (β2-M). (B) Cellular lysates were prepared from equal numbers of uninfected A549 and BJAB cells or Ad5dl309-infected BJAB cells at 3 and 174 days postinfection and analyzed by immunoblotting for CAR or actin as a loading control. (C) BJAB cells were infected with Ad5dl309 at 100 PFU/cell. Cellular RNA was extracted at various days after infection and analyzed by reverse transcription followed by PCR to detect mRNA for CAR and β₂-microglobulin. (D) BJAB cells were infected with Ad5dl309 at 100 PFU/cell. The persistently infected cells were maintained in culture medium for 75 days and then superinfected with AdCMV-GFP. GFP-positive cells were detected by flow cytometry. dpi, days postinfection.