Downstream of tyrosine kinase/docking protein (Dok)6 colocalises with tropomyosin-related kinase (Trk)C and binds to TrkC receptor in E18.5 mouse brains. (a) Specificity tests for TrkC (R&D Systems) and Dok6 (Abcam) antibodies. Human embryonic kidney (HEK)293 cells were transfected with V5-TrkC and Flag-Dok6 separately. Cell lysates and homogenates of E18.5 mouse brains were then detected by western blotting with anti-TrkC and anti-Dok6 antibodies separately, using anti-V5 and anti-Flag antibodies as positive controls (panels 1 and 2). HEK293 cells were transfected with V5-TrkA, V5-TrkB and V5-TrkC separately. Cell lysates were then detected by western blotting with anti-TrkC and anti-V5 antibodies (panels 3 and 4). (b) TrkC colocalises with Dok6 in E18.5 mouse primary cortex cells. Immunostaining in cortex cells with anti-TrkC and anti-Dok6 antibodies. Cells were counterstained with 4',6-diamidino-2-phenylindole (DAPI) to show nuclei. Scale bar: 10 μm. (c) Dok6 binds to the TrkC receptor in mouse brain. Homogenates of E18.5 mouse brains were immunoprecipitated with anti-Dok6 and IgG antibodies; the bound proteins were then analysed by western blotting with anti-TrkC, anti-Dok6 and anti-pTyr antibodies.
Li et al. BMC Biology 2010 8:86 doi:10.1186/1741-7007-8-86