Figure 2.

 Spliceostatin A (SSA) suppressed vascular endothelial growth factor (VEGF) production. (a) HeLa cells were treated with 200 nM SSA or inactive acetylated‐SSA (Ac‐SSA) for the indicated times, and RT‐PCR analysis was performed using primers that detect both pro‐angiogenic VEGF₁₆₅ (∼200‐bp) and anti‐angiogenic VEGF₁₆₅b, (∼150‐bp). (b) HeLa cells were treated with 200 nM SSA or FR901464 (FR) for 3 or 12 h, and then western blot analysis using anti‐VEGF antibody was performed. The GAPDH was a loading control. (c) HeLa cells were treated with vehicle (0.1% methanol; MeOH), 200 nM FR901464, 200 nM SSA or Ac‐SSA for the indicated times. The levels of secreted VEGF protein in the medium were determined by ELISA and were normalized by cell numbers in the same dishes. The results are expressed as percentages of the control cells treated with vehicle as the means ± SD of three independent experiments, each performed in duplicate. ***P < 0.001; *P < 0.05.