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For immunohistochemical staining, we incubated the sections with primary antibodies overnight at 4 C and used a horseradish peroxidase-streptavidin detection system (Dako) to detect immunoactivity followed by counter-staining with hematoxylin (Sigma-Aldrich)

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The WT or mutant constructs were transiently transfected using Lipofectamine 2000 transfection reagent as previous described 25

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are co-inventors of a patent application related to this work

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