NESH knockdown reduces synapse formation and affects the postsynaptic apparatus.

<p>(A) Hippocampal neurons were transfected with the control (scrambled siRNA) or NESH siRNA at 10–12 DIV and stained with anti-VAMP2 antibody, anti-GluR1 antibody or Alexa Fluor 594-conjugated phalloidin at 16–18 DIV. GFP was co-transfected with siRNAs to visualize dendritic spines. (B) Synapse formation per µm was analyzed in NESH knockdown neurons and compared with control (n = 15 for control; n = 19 for NESH siRNA). (C) Numbers of GluR1 cluster per µm on spines (n = 17 for control; n = 15 for NESH siRNA). (D) F-actin fluorescence intensity ratios (spine vs. shaft; n = 15 for control and NESH siRNA). Data are presented as means ± SEM. *p<0.05, **p<0.01.