Mass spectrometry analysis identifies SOX10 phosphorylation sites.

<p>A. Workflow schematic of SOX10 protein analysis by mass spectrometry. 501mel cells were treated with MG132 proteasomal inhibitor before scraping cells and performing immunoprecipitation (IP) using SOX10 antibody to isolate protein. The eluted proteins were separated by SDS-page, followed by staining and removal of bands corresponding to 55kD, 75kD and 100kD. All three gel bands were subjected to destaining, in-gel digestion and extraction before running LC-MS/MS. B. Portions of IP samples were separated on SDS-page gel, followed by transfer onto PVDF membrane and Western blotting to confirm SOX10 isolation in the eluted samples being used for mass spectrometry. C. The three phosphorylation sites selected for mutation and characterization are shown in the context of full length SOX10 (Genbank ID NM_006941).