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. 2014 Oct 28;111(45):16154–16159. doi: 10.1073/pnas.1401483111

Fig. 1.

Fig. 1.

Identification of the Kif5C complex in mouse hippocampus. (A) Strategy used to analyze Kif5C-specific protein cargos in the mouse hippocampus. The schematic diagram shows the workflow for profiling Kif5C-transported proteins by proteomics. The processes include brain dissection, hippocampal tissue extraction and homogenization, Kif5C IP, complex separation by SDS/PAGE, isolation of specific bands followed by in-gel tryptic digestion, and LC-MS/MS analysis. (B) IP of Kif5C from mouse hippocampus. Kif5C complexes were immunoprecipitated from hippocampus using anti Kif5C antibody. (Upper) Protein complexes were then separated on SDS/PAGE and stained with silver. (Lower) Kif5C also was detected by Western blot analysis. Immunoprecipitation using CREB antibody and beads alone served as controls for IP. (C) Co-IP and Western blot analysis of Kif5C cargo proteins (PURα, DIC1, and GluR1) in mouse hippocampus. Total extract (input), IP, post-IP supernatant, beads-alone IP, and post-IP supernatant are shown. Arrows in B indicate the positions of Kif5C and antibody.