Journal: Cancers

Article Title: WNT5A-Induced Activation of the Protein Kinase C Substrate MARCKS Is Required for Melanoma Cell Invasion

doi: 10.3390/cancers12020346

Figure Lengend Snippet: MARCKS phosphorylation-specific peptide MANS blocks WNT5A-induced melanoma cell invasion. Western blotting and transwell invasion assays were performed to evaluate the specific role of MARCKS phosphorylation in WNT5A-promoted melanoma cell invasion. ( A ) Western blots showing the expression of total MARCKS and levels of pMARCKS Ser-159/163 in 100 µM RNS- or MANS peptide-treated A2058 melanoma cells. Representative blots of five separate experiments are shown here. ( B,C ) The graph represents the densitometry analysis of ( B ) MARCKS and ( C ) pMARCKS Ser-159/163 levels normalized against total MARCKS levels. The results (n = 5) are presented as the means ± S.E.M.; ***, p < 0.001. ( D ) Effect of the direct inhibition of pMARCKS on WNT5A-induced melanoma cell invasion was evaluated by transwell cell invasion assay as mentioned in the materials and methods section. ( D ) Graph showing the relative melanoma cell invasion of rWNT5A unstimulated/stimulated and RNS peptide- or MANS peptide-treated A2058 melanoma cells. The results (n = 4) are presented as the means ± S.E.M.; *, p < 0.05, and ***, p < 0.001.

Article Snippet: The MANS and RNS peptides were custom synthesized by CASLO ApS (Lyngby, Denmark).

Techniques: Phospho-proteomics, Western Blot, Expressing, Inhibition, Invasion Assay