Fig. 5

Overexpression of nestin does not rescue the neurite outgrowth defect in DLK-depleted cells. (A) Control (pLKO.1) and shDLK#2/DLK-depleted Neuro-2a cells were transfected with plasmids expressing either EGFP (control) or wild-type nestin and grown in proliferation medium. At 48 h after transfection, the cytoskeleton was prepared and processed for western blot analysis using antibodies against nestin and vimentin. Bands of interest were excised from different blots and grouped together. Full-length blots are presented in Supplementary File 4. (B) Neurite length (mm) per cell-body cluster was measured every 4 h in control and shDLK#2/DLK-depleted cells that were differentiated from RA for 76 h two days after transfection with EGFP or nestin. Data on graph represent the individual values obtained from three independent experiments at each time point with means connected. Multiple unpaired t tests were used for statistical analysis. ns, not significant