cAMP levels and forskolin effects are different in 2D

<p>- <b>and 3D-cultured cells.</b> (A) cAMP levels are higher in 3D-cultured cells compared to 2D-cultured cells. Cells were grown in 2D or 3D cultures for 24 h and analyzed for cAMP. Each bar represents the mean value ± S.D. of three independent experiments. (B) cAMP/TGFβ-induced changes in RNA levels are translated into changes in protein levels. MDA-MB-231 cells were incubated with forskolin or TGFβ1 or both or mock-treated for 24 h, lysed and analyzed for protein levels of TIMP-1, Cox-2 and PAI-1 by the Western blot technique. The level of the secretory protein TIMP-1 was measured in the medium (MD) in which cells were grown, Cox-2 protein expression was analyzed in plasma membrane (PM) extracts, PAI-1 levels were determined in both MD and PM. To check for equal loading gels were stained with Coomassie Blue.