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Journal: eNeuro
Article Title: Loss of SynDIG1 Reduces Excitatory Synapse Maturation But Not Formation In Vivo
doi: 10.1523/ENEURO.0130-16.2016
Figure Lengend Snippet: Synapse composition is unaltered in SynDIG1-deficient synapses. A , Representative immunoblots of biochemical fractions isolated from WT and SynDIG1 β-gal homozygous mutant P14 mouse brain tissue showing levels of GluA1, GluA2, GluN1, GluN2B, PSD-93, PSD-95, synaptophysin (Synapto), SynDIG1, SynDIG4, and PICK-1 present in the S1-, P2-, Syn-, and PSD-enriched fractions. Loading controls are provided by β-actin and β-tubulin immunoreactivity. B–D , Graphs depict the ratio of SynDIG1 β-gal homozygous mutant protein relative to WT levels of AMPA receptor subunits ( B ), NMDA receptor subunits ( C ), and PSD-93 and PSD-95 ( D ) in the PSD-enriched fractions. Data are the average of three independent biochemical fractionation experiments; each experiment used four to six mouse brains of each genotype. Error bars represent ±SEM.
Article Snippet: Membranes were blotted with the following primary antibodies: mouse antibodies against PSD-95 [catalog #75-028, NeuroMab (RRID:AB_2292909)], synaptophysin [catalog #101011, Synaptic Systems (RRID:AB_887824)], SynDIG1 [catalog #75-251, NeuroMab (RRID:AB_10999753)], GluA2 [catalog #75-002, NeuroMab (RRID:AB_2232661)], PSD-93 [catalog #75-057, NeuroMab (RRID:AB_2277296)], β-tubulin [catalog #05-661, Millipore (RRID:AB_309885)], GluN1 [catalog #556308, BD Biosciences (RRID:AB_396353)], and Pick1 [catalog #73-040, NeuroMab (RRID:AB_10672986)];
Techniques: Western Blot, Isolation, Mutagenesis, Fractionation