Effect of F-actin stabilization on NESH mobility in dendritic spines.

<p>FRAP analysis was performed to examine the effect of F-actin stabilization on NESH mobility. Hippocampal neurons at 10–12 DIV were transfected with GFP-NESH, GFP, GFP-actin, GFP-PSD95 or GFP-Homer1c, and subjected to FRAP analysis at 16–18 DIV. (A, F) Recovery curves of GFP-NESH in non-treated (control) and jasplakinolide-treated neurons (mobile fractions: 43.9±2.4% at 400 s for control, 13.6±2.8% at 400 s for jasplakinolide). (B, F) Recovery curves of GFP (mobile fractions: 92.7±8.0% at 400 s for control, 83.3±1.3% at 400 s for jasplakinolide). (C, F) Recovery curves of GFP-actin (mobile fractions: 98.6±8.7% at 400 s for control, 0.2±0.1% at 400 s for jasplakinolide). (D–F) Recovery curves and mobile fractions of the scaffolding proteins, PSD95 and Homer1c (control: 16.7±5.2% at 400 s for GFP-PSD95, 14.8±6.6% at 400 s for GFP-Homer1c, jasplakinolide: 12.0±2.7% at 400 s for GFP-PSD95, 7.0±4.2% at 400 s for GFP-Homer1c). (F) Analysis of the mobile/immobile fractions from data obtained in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034514#pone-0034514-g003" target="_blank">Fig. 3A–E</a> (N = 15 for each condition, data from three to five independent experiments). Data are presented as means ± SEM. **p<0.01.