CREB Inhibits AP-2α Expression to Regulate the Malignant Phenotype of Melanoma
Figure 2
Inhibition of pCREB results in increased AP-2α expression.
(A) pCREB was analyzed in non-metastatic and metastatic cell lines by Western Blot. As confirmed by densitometry, pCREB in metastatic cell lines is approximately 5-fold higher as compared to non-metastatic cell lines. α-Actin is used as a loading control. (B) SB2 and C8161-c9 cell lines were immunofluorescently stained for detection of pCREB (Red). Colocalization with DAPI (Blue) and pCREB reveal high pCREB intensity in the nuclei of C8161-c9 cells (low AP-2α) while low level of pCREB is mostly undetectable in the nuclei of SB2 cells (high AP-2α). (C) Expression of AP-2α was detected in A375SM and C8161-c9 cell lines after incubation with the pCREB inhibitor, H-89. Complete restoration of AP-2α expression was observed in both cell lines after treatment with H-89 at 10 mM. Lamin was used as a loading control.