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Image Search Results
Journal: Journal of Neurochemistry
Article Title: α-Synuclein forms a complex with transcription factor Elk-1
doi: 10.1046/j.1471-4159.2001.00232.x
Figure Lengend Snippet: Fig. 5 Elk-1 does not bind to a-synuclein, but ERK-2 binds to a-synuclein. (A) Neither normal nor phosphorylated Elk-1 binds to a-synuclein in vitro. Bacterial expressed Elk-1 was phosphorylated by ERK-2. Neither non-phosphorylated (Elk-1) nor phosphorylated (p-Elk-1) forms bound to GST-a-synuclein. (B) a-Synuclein binds to ERK-2, and Elk-1 binds to ERK-2 in vitro. Bacterial expressed a-synuclein and Elk-1 were mixed with GST-ERK-2, and pull- down assay was performed using glu- tathione sepharose 4B. Washed beads were subjected to SDS-PAGE and immuno- blotted with S1 or anti-Elk-1 antibodies. GST and GST-ERK-2 were stained with anti-Elk-1 antibody non-speci®cally. (C) ERK-2 colocalizes at cytoplasmic granules. OS-3 cells were cotransfected with pcDNA3.1V5His ERK-2, pcDNA4mycHis Elk-1 and pCI a-synuclein. Cells were ®xed and stained with S1 and anti-V5 antibodies. Secondary antibodies were Alexa 546 anti- rabbit and Alexa 488 anti-mouse antibodies. Bar 10 mm.
Article Snippet: 6-His-tagged protein was puri®ed using
Techniques: In Vitro, Pull Down Assay, SDS Page, Staining