Figure 2

Comparison of protein production rate of the F-bacmid and original bacmid in insect and mammalian cells. Medium was harvested from insect and mammalian cells at 96 and 48 hours after virus addition, respectively. Equal amount of total protein was loaded into immunoblots. (A) The VEGF-D^ΔNΔC production in insect cells analyzed by ELISA. (B) Immunoblot of VEGF-D^ΔNΔC production in insect cells. Lanes 1–4 represent medium samples of FG-VEGF-D^ΔNΔC produced by conventional bacmid system and the three F-bacmid produced VEGF-D^ΔNΔC. Lanes 5–8 show cell lysates from the same samples. Lane 9 is the positive control for VEGF-D. (C) Lanes 1–4 are the medium samples from two avidin controls produced by conventional bacmid system (lanes 1 and 2) compared to two F-bacmid produced avidins (lanes 3–4). Lanes 5–8 are cell lysates from the same samples. Lane 9 is positive control for avidin. (D) ELISA results for theVEGF-D^ΔNΔC production in mammalian cells. (E) An immunoblot analysis of avidin production in mammalian cells. Lanes 1 and 2 represent HepG2 medium samples of avidin control and F-bacmid avidin, respectively. Lanes 3–4 show the medium samples from BT4C cells, and lanes 5 and 6 from avidin produced in SKOV 3 cells. Lane 7 is the positive control for avidin. (M) Marker (SeeBlue^® PreStained Standard, Invitrogen).