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. 2014 Jul 3;4:5546. doi: 10.1038/srep05546

Figure 4. Stimulation of cAMP/CREB signaling pathway by CG500354 and mimetic substances.

Figure 4

(A) Measurement of the extracellular cAMP level was performed with a Cyclic AMP EIA Kit. The supernatant from the CG500354-treated GBM cells showed a significantly higher level of cAMP than that from the DMSO-treated GBM cells. (B) Western blot analysis shows that the expression of phosphorylated PKA and CREB was strongly induced by CG500354 treatment. The full-length blots are included in the supplementary information (Suppl. Fig. S7). (C) Both mimetic substances (Forskolin and Rolipram) and si-PDE4D also increased the expression of phosphorylated CREB. The full-length blots are included in the supplementary information (Suppl. Fig. S8). (D) Quantitative RT-PCR analysis reveals that both mimetic substances induced p21 expression and only Rolipram also increased p27 expression. (E–F) GBM-derived cells treated with CG500354, Forskolin or Rolipram were stained with a p53 antibody. The numbers of p53-expressing cells (green) in the three groups were all equivalently high. Nuclei (blue) were counterstained with DAPI. Scale bar = 0.5 mm.