Homer1

For the analysis of synaptic marker expression, hippocampi from WT and Myo9a^+/- mice were dissected and homogenized in RIPA buffer (20 mM Tris, 150 mM NaCl, 1 mM EDTA, 1% NP40, 1% Triton X-100, protease inhibitors, pH 7.5). Ten micrograms of protein from each sample were loaded onto an acrylamide gel for western blotting_.Primary antibodies directed against the following proteins were used: Myo9a (Tü 76, WB 1:1000; Tü 78, IP 10 μg/ml; Chieregatti et al., 1998 (link); Hanley et al., 2010 (link)), Myo5a (AbCAM, 1:1000), GluA1 (Chemicon, 1:1000), GluA2 (Chemicon, 1:1000), GluA2/3 (1:2000, gift from C. Gotti), GluK2 (Prestige, 1:1000), Homer1 (Santa Cruz, 1:500), PSD95 (Neuromab, 1:20000), Synaptophysin1 (SySy, 1:5000), GAPDH (Santa Cruz, 1:1000), N-cadherin (NCAD) (AbCAM, 1:1000), PICK1 (Neuromab, 1:1000), GRIP1 (BD Transduction Laboratories, 1:2000), Tubulin (Sigma, 1:50000), Transferrin receptor (Invitrogen, 1:1000), VGAT (SySy, 1:1000), and VGLUT (SySy, 1:2000). The secondary antibodies, horseradish peroxidase- (Sigma) or IR Dyes- (LI-COR) conjugated antibodies, were used for western blots. Samples were separated using SDS-PAGE, and western blots then visualized with the Pierce ECL-Detection Kit or an Odyssey Infrared Imager (LI-COR).

Molecular weight marker, reagents and nitrocellulose membranes for SDS/PAGE were acquired from Bio‐Rad (Mississauga, ON, USA) and Millipore Sigma (GE10600002, Burlington, MA, USA). Antibodies against soluble amyloid precursor protein β (sAPPβ; 1 : 500; BioLegend, San Diego, CA, USA, cat# SIG‐39138), sAPPα (1 : 500, BioLegend cat#SIG39139), APP (1 : 500 BioLegend, cat#SIG039152), BACE1 (1 : 500; Cell Signaling, Whitby, Canada, cat#5606P), synaptophysin, acquired from Cell Signaling Technology (concentrations cat#5461); PSD‐95, acquired from Santa Cruz Biotechnology (concentrations cat# sc‐32290); Homer1, acquired from Santa Cruz Biotechnology (concentrations cat#sc‐136358). Horseradish peroxidase anti‐mouse and anti‐rabbit secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, West Grove, PA, USA (Donkey anti‐rabbit IgG (H + L) 711‐035‐152, Goat anti‐mouse IgG (H + L) 115‐035‐003 Jackson ImmunoResearch) and Western Lightning Plus Enhanced Chemiluminescence from Perkin Elmer (Guelph, Canada, cat#105001EA).