Figure 7.

Down-regulation of TβRII causes dermal DF and HDMEC motility to become TGFβ3 insensitive just like the epidermal cells. DFs and HDMECs were infected with various dilutions (vol/vol) of the lentivirus (FG-12) carrying an siRNA against TβRII. After 48 h, the cells were first analyzed for down-regulation of the endogenous TβRII before being subjected to migration assays. (A) 50% dilution of the original virus stock decreases TβRII expression in DFs (a, lane 6) to the level similar to HKs (a, lane 3 vs. lane 1). (B) The same dilution of the virus stock decreases the TβRII expression in HDMECs (c, lane 6) to the similar level in HKs (c, lane 3 vs. lane 1). (C) Coexpressed GFP and FACS analyses show no significant reduction in gene transduction efficiency in HDMECs (a–c) or DFs (e–g) unless the virus stock is diluted to 25% or further (d and h). (D and E) DF and HDMEC cells 48 h after infection with 50% of the original virus stocks of vector alone, lac-Z–siRNA, and TβRII-siRNA were subjected to colloidal gold migration assays in the absence or presence of plasma, serum, and plasma plus TGFβ3. MIs are shown. These migration experiments were repeated three times. Error bars represent SEM.