Fig. 3.

Gossypol decreases vascular endothelial growth factor (VEGF) mRNA stability and thereby its protein translation. a Quantitative RT-PCR for VEGF mRNA expression in MCF-7 and MDA-MB-468 cells treated with 10 μM gossypol for different time periods. b Pulse-chase and quantitative RT-PCR for the stability of VEGF mRNA in MDA-MB-468 cells treated with gossypol. c MDA-MB-468 cells were treated with or without 10 μM gossypol for 8 h, and then their cytoplasmic lysates were fractionated on a sucrose gradient. RNA extracted from each of the fractions was subjected to quantitative RT-PCR for quantitative analysis of the distribution of VEGF mRNA. Data represent percentage of the total amount of corresponding mRNA for each fraction. d Western blot for expression of MDM2 and VEGF in MDA-MB-468 cells with MDM2 knockdown treated with indicated doses of gossypol for 24 h