Decreased Notch pathway activity in isolated MAFs.

A. Characterization of isolated MAFs by morphology (s) and IF (color images), and comparison with normal human skin fibroblasts and melanoma cells. A single scale bar in a panel of pictures is representative for all pictures. B. Quantification of gene expression of the Notch pathway components in MAF vs. FF2441 by PCR Array. The Notch1, Notch4, Dll1, Hes-1 and Hey-1 genes were significantly downregulated (<2-fold) in MAFs (* P<0.01). The results are mean ± SD of three independent experiments. C. The decreased protein expression of Notch1, Notch4, Dll1, Hes-1 and Hey-1 in MAFs vs. FF2441 was confirmed by immunoblot. Blots were cropped from different gels. β-actin was used as a loading control. D. The decreased protein expression of Notch1 and Hes-1 in MAFs vs. NHDF was confirmed by immunoblot. Blots were cropped from different gels. β-actin was used as a loading control. For C and D, Uncropped images of immunoblot are available in S1 File. Relative levels of proteins from three independent Western blots are shown [proteins expressed in normal skin fibroblasts [FF2441 and NHDF, respectively) are set as “1”].