Knockdown of Nischarin expression promotes neurite outgrowth in PC-12 cells.

<p><b>(A)</b> Fluorescence images of PC-12 cells five days after infection with Nis-shRNA-3 or control-shRNA lentivirus. Four days after the infection, PC-12 cells were exposed to NGF for 24 h. Scale bar: 20 μm. <b>(B, C)</b> Quantification of neurite outgrowth five days after lentivirus infection. The neurites analyzed are processes from eGFP-positive cells under a confocal microscope (n = 3, ~200 cells measured). Suppressing Nischarin expression increases the mean length of the longest neurite and the total length of neurites per cell. Asterisks indicate significant differences compared with control-shRNA cells (*<i>p</i><0.05, **<i>p</i><0.01). Data presented are the mean ± SEM. <b>(D)</b> Images from time-lapse recordings of PC-12 cells after infection by Nis-shRNA-3 or control-shRNA lentivirus. Time-lapse images were taken at three days post-infection in 45 min intervals to monitor the velocity of neurite extension in PC-12 cells. Scale bar: 20 μm. <b>(E)</b> Quantification of the average velocity of neurite extension.