Fig. 2. Lacking Arhgef4 decreased the number of dendritic protrusions but increased the size of PSD-95 particles. (A) Hippocampal neurons isolated from WT and Homo mice were cultured. Neurons were infected with GFP Sindbis viruses and immunostained with monoclonal PSD-95 antibodies. Scale bar: 10 μm. (B) The selected dendrites were straightened and enlarged. Scale bar: 10 μm. (C) The size of PSD-95 particles was increased in the neurons of Arhgef4 knockout mice (Arhgef4+/+: 100.0%±4.93%, N=15 dendrites; Arhgef4-/-: 117.3%±5.98%, N=14 dendrites, two-tailed unpaired t-tests: *p<0.05). (D) The number of dendritic protrusions per 10 μm was decreased in the neurons of Arhgef4 knockout mice (Arhgef4+/+: 8.2±0.86, N=15 dendrites; Arhgef4-/-: 4.3±0.39, N=14 dendrites, two-tailed unpaired t-tests: **p<0.01). (E) Ablation of Arhgef4 gene did not change synaptic protein expression levels, including PSD-95, in the brain. Brain lysates (25 μg of lysate for PSD-95, Homer-1, GluA1, and GluN1, 3 μg of lysate for synaptophysin, α-tubulin) from WT, Hetero, and Homo mice were loaded and subjected to western blotting analysis.
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