Downstream of tyrosine kinase/docking protein (Dok)6 plays a role in neurite outgrowth in E18.5 mouse cortex primary neurons. (a) Knockdown efficiency tests of Dok6i construct and nonsense control construct. Human embryonic kidney (HEK)293 cells were cotransfected with Flag-Dok6 and Dok6i or Flag-Dok6 and nonsense control constructs separately. The cell lysates were then analysed by western blotting with anti-Flag antibody. (b) E18.5 mouse cortex primary cells were transfected with Dok6i and nonsense control constructs, both of which were green fluorescence positive in cells. An immunofluorescence test was performed 48 h later with anti-microtubule-associated protein 2 (MAP2; red). and photographs of cells that were positive for both red and green fluorescence were taken (white arrow). Scale bars: panel a 50 μm, panel b 200 μm, panel c-h 30 μm. (c) Quantification of neurite outgrowth in cortex neurons. In all, 10 random fields of cells from each experimental group were photographed. Neurite length was quantified by measuring the longest neurite processes of cells that were positive for both green and red fluorescence by using NIS-Elements software. *P value < 0.05 by Student t test.
Li et al. BMC Biology 2010 8:86 doi:10.1186/1741-7007-8-86