Fig. 4: SH3RF3 interacts with presynaptic proteins.
From: Sh3rf3 Deficiency drives autism-like behaviors via presynaptic dysfunction in mice
(A) Schematic representation of the construction strategy for Sh3rf3-3 × flag mice (Top), Enrichment of adult WT and Sh3rf3-3 × flag mouse PFC tissue samples using FLAG beads (Bottom). (B) Analysis of IP-MS data. Dot graph showing the binding probabilities of prey proteins, with a dashed line indicating the threshold of MaxP = 0.5 (Left-top). Bar graph showing the GO-BP enrichment terms of repeatedly detected prey proteins with MaxP > 0.5 (Left-bottom). Network diagram of some significantly enriched GO-BP terms and proteins in them (Right). AvgP, average binding probability; MaxP, maximum binding probability. (C) Exogenous immunoprecipitation analysis demonstrated that SH3RF3 is capable of interacting with ERC2, Synapsin1, RIM1 and Liprin α3. (D) Interaction between SH3RF3 and presynaptic proteins ERC2, Synapsin1, RIM1 and liprin α3 was detected in mouse PFC. (E) Subcellular distribution. Mouse brain homogenates were subjected to subcellular fractionation, and an aliquot of each fraction (10 μg) was analyzed by western blotting with the indicated antibodies. The results are representative of three independent experiments. Hom, homogenate; S1, crude synaptosomal fraction; P2, crude membrane fraction; P2C, synaptosomal fraction; CSM, crude synaptic membrane fraction; CSV, crude SV fraction; SM3, synaptosomal membrane; PSD, postsynaptic density fraction; S, 1% (w/v) Triton X-100-soluble fraction of PSD; P, 1% Triton X-100-insoluble fraction of PSD. (F) Illustrative diagram of SH3RF3 interacting with presynaptic proteins.
