Expression profile of NESH in brain and cellular localization in hippocampal neurons.

<p>(A) During development, the expression of NESH was examined by immunoblot analysis in postnatal whole brains (from P7 to adult). (B) Adult rat brains were dissected into subregions, and the lysates were subjected to immunoblot analysis using anti-NESH antibody. SPIN90 and Homer1c were used as controls for PSD proteins; β-actin served as a loading control. (C) Hippocampal neuron lysates were fractionated to crude synaptosomal (P2) and cytosolic fractions (S2), and the equal amount of NESH in each fraction was examined by immunoblotting. (D) Cellular localization of NESH was examined in primary cultured hippocampal neuron at 19 DIV. Mature hippocampal neurons were double-stained with antibodies against NESH, SPIN90 or PSD95 and with Alexa Fluor 594-conjugated phalloidin. Boxed regions were magnified for better imaging of co-localization. Arrows indicate co-localized regions of the images. The scale bar represents 20 µm.