Methods: Using mouse endothelial stem cells (MEES), we examined the global proteomic effects of the loss of each CCM gene expression. Label-free differential protein expression analysis using multidimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) was applied to examine the expressed proteomic profile for each knockdown cell-line (CCM1, CCM2, and CCM3) compared to mock shRNA and no shRNA control cell-lines. Those differentially expressed proteins were identified (ANOVA; p<0.05) and principle component analysis (PCA) as well as cluster analysis were used to analyze identified proteins.
Results: 122 proteins were found to be differentially expressed amongst the five cell-lines. PCA and cluster analysis results demonstrate the effects of individual CCM knockdown and suggests a unique proteomic profile for each cell line. In each of the three CCM knockdown cell-lines, overexpression of cytoskeletal proteins is the most common.
Conclusions: The results suggest that these CCM genes play essential role in cytoskeletal development and that proteomic analysis is proven to be a useful tool for future examination of CCM-related signaling pathways.
Keywords: Cell biology, Cell culture, Central nervous system/peripheral nervous system and Neuroscience
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