Fig. 2.

Antiproliferative effects resveratrol in DNA synthesis and cyclin D1 downregulation. (A) B103 cells were culture on a round cover slides in 24 well dishes until 80% confluence in 10% FBS. The cells were then treated in the absence and presence of 15 µM of resveratrol. After 46 h resveratrol treatment, cells were incubated with 10 µM BrdU for 2 h before fixation and subsequent immunostaining with an anti-BrdU antibody (Arrow shows BrdU positive cells). (B) The percentages of stained cells were counted in three independent random areas. Results are means±SE and representatives of three independent experiments are shown (n=3, ^*p<0.05). (C) B103 cells were cultured in 60-mm culture dishes to near 80% confluence in 10% FBS. They were treated with 0 to 20 µM of resveratrol for 48 h. Equal volumes of whole-cell extracts containing 40 µg of protein were separated and electrophoretically blotted. Cyclin D1 was detected via immunoblot analysis. β-actin was used as a loading control. (D) The intensities of the cyclin D1 bands were determined by densitometric scanning and analyzed by Bio-Profil software and the expression levels were normalized to β-actin. Results are mean±S.E. and representatives of three independent experiments are shown (n=3, ^*p<0.05, ^**p<0.01).