|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 5. Involvement of PKC in TGFß1/ROS-dependent expression of 
SMA. (A) Subconfluent fibroblasts in CMHDF were preincubated with the PKC inhibitors Ro 31-8220 and Ro 32-0432 for 1 hour before treatment with 10 ng/ml rTGFß1 in combination with each inhibitor. Expression of SMA was detected by western blots. The densitometric analysis describes protein expression as fold increase over control, which was set at 1.0. The data represent the mean ± s.e.m. of three independent experiments. (B) Time-course analysis for activation of PKC was performed using subconfluent HDFs either untreated (C) or treated with 10 ng/ml rTGFß1 CMHDF for the indicated time periods. The cell lysates were subjected to western blot analysis for phospho-PKC. The image is representative of two independent experiments. The densitometric analysis represents fold increase over control (C) which was set at 1.0. (C) Subconfluent HDFs were preincubated with the antioxidants as described prior to addition of 10 ng/ml rTGFß1 for a further 1 minute. Western blot analysis was performed for phospho-PKC and total PKC. Total PKC bands indicate the loading control. Data shown are representative of three independent experiments. (D) Subconfluent HDFs were preincubated with PKC inhibitor RO 31-8220 for 1 hour (closed circles) before treatment with rTGFß1 in CMHDF or 1 mM H2O2 for the indicated time. Increase of DCF fluorescence was followed over 15 minutes versus untreated controls (open circle). The experiments were performed in duplicate. Arrows indicate addition of rTGFß1 or H2O2.
|
