FIGURE

Fig. 6

ID

ZDB-FIG-220802-60

Publication

Chen et al., 2022 - Autism-Risk Gene necab2 Regulates Psychomotor and Social Behavior as a Neuronal Modulator of mGluR1 Signaling

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Fig. 6

NECAB2 interacts with mGluR1 in vitro. (A) Overview of the protein interaction network derived from the Necab2 co-immunoprecipitation and subsequent mass spectrometry in the necab2^+/+ and necab2^–/– larvae. (B) REACTOME analysis revealed the significant biological processes detected in the necab2^+/+ larvae compared to the necab2^–/– larvae. (C) Gene Ontology (GO) analysis revealed the significant biological processes detected in the necab2^+/+ larvae compared to the necab2^–/– larvae. (D–D″′) NECAB2 interacts with mGluR1 through the NHR domain uncovered by co-immunoprecipitation analysis. The HEK293 cells were transiently transfected with mGluR1-mCherry plus NECAB2-Myc (D), EGFP-EF (D′), EGFP-NHR (D″), or EGFP-ABM (D″′) for lane 1 and lane 3 respectively, and indicated vectors plus mCherry-vector for lane 2 and processed for immunoprecipitation using mouse anti-Myc antibody (D) or anti-EGFP antibody (D–D″′), with normal mouse IgG for negative control in lane3. The crude extracts (Input) and immunoprecipitations (IP) were analyzed by SDS-PAGE and immunoblotted using the rabbit anti-mCherry antibody (D–D″′), mouse anti-Myc antibody (D) or mouse anti-EGFP antibody (D′–D″′). (E–E″′) NECAB2 has self-interaction through either NHR or ABM domain uncovered by co-immunoprecipitation analysis. The HEK293 cells were transiently transfected with NECAB2-Flag plus EGFP-NECAB2 (E), NECAB2-Flag plus EGFP-EF (E′), NECAB2-Flag plus EGFP-NHR (E″), or NECAB2-Flag plus EGFP-ABM (E″′) for lane 1 and lane 3, and the indicated vectors plus EGFP-vector for lane 2 and processed for immunoprecipitation using mouse anti-Flag antibody, with normal mouse IgG for negative control in lane 3. The crude extracts (Input) and immunoprecipitations (IP) were analyzed by SDS-PAGE and immunoblotted using mouse anti-EGFP antibody (E–E″′) or mouse anti-Flag antibody (E–E″′). (F–F″) Co-immunoprecipitation analysis of NHR and ABM domains showed self-interaction but no cross-interaction of the two domains. The HEK293 cells were transiently transfected with Flag-NHR plus EGFP-NHR (F), Flag-ABM plus EGFP-ABM (F′), and Flag-ABM plus EGFP-NHR (F″) for lane 1 and lane 3, and indicated vectors plus EGFP-vector for lane 2 and processed for immunoprecipitation using the mouse anti-Flag antibody, with normal mouse IgG for negative control in lane3. The crude extracts (Input) and immunoprecipitations (IP) were analyzed by SDS-PAGE and immunoblotted mouse anti-Flag antibody (F–F″) or mouse anti-EGFP antibody (F–F″).

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	necab2^s4301/s4301
Observed In:	G protein-coupled receptor signaling pathway G protein-coupled serotonin receptor signaling pathway G-protein beta/gamma-subunit complex binding phospholipase C-activating G protein-coupled receptor signaling pathway
Stage:	Day 5

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Front. Mol. Neurosci.