Journal: eLife

Article Title: The anterior cingulate cortex and its role in controlling contextual fear memory to predatory threats

doi: 10.7554/eLife.67007

Figure Lengend Snippet:

Article Snippet: For the pharmacogenetic inhibition, ACA was injected bilaterally with 150 nl of AAV5-hSyn-HA-hM4D(Gi)-IRES-mCitrine (Dr. Bryan Roth; Addgene plasmid #50464) or AAV5-hSyn-eGFP (titer≥7×1012 vg/ml; Addgene viral prep #50465-AAV5) as control.

Techniques: Transfection, Construct, Virus, Plasmid Preparation, Software

Journal: medRxiv

Article Title: Identification and molecular characterization of missense mutations in orphan GPCR GPR61 occurring in severe obesity

doi: 10.1101/2024.10.21.24315834

Figure Lengend Snippet: RMSD of carbon alpha atoms from the three simulations of the GPR61 WT ( A ) and GPR61 R236C ( B ).

Article Snippet: HiBiT-GPR61 plasmid DNA was generated with Gibson cloning using GPR61-Tango plasmid DNA (#66366 Addgene, deposited by Bryan Roth) and HiBiT-FZD 6 plasmid DNA as templates.

Techniques:

Journal: medRxiv

Article Title: Identification and molecular characterization of missense mutations in orphan GPCR GPR61 occurring in severe obesity

doi: 10.1101/2024.10.21.24315834

Figure Lengend Snippet: The same procedure was used for GPR61, MC 4 R and GPR141.

Article Snippet: HiBiT-GPR61 plasmid DNA was generated with Gibson cloning using GPR61-Tango plasmid DNA (#66366 Addgene, deposited by Bryan Roth) and HiBiT-FZD 6 plasmid DNA as templates.

Techniques:

Journal: medRxiv

Article Title: Identification and molecular characterization of missense mutations in orphan GPCR GPR61 occurring in severe obesity

doi: 10.1101/2024.10.21.24315834

Figure Lengend Snippet: A. Mutational landscape of GPR61 in normal population and then the same mutations in severe obesity patients. B. GPR61 mutations mapped onto a 2D model of GPR61.

Article Snippet: HiBiT-GPR61 plasmid DNA was generated with Gibson cloning using GPR61-Tango plasmid DNA (#66366 Addgene, deposited by Bryan Roth) and HiBiT-FZD 6 plasmid DNA as templates.

Techniques:

Journal: medRxiv

Article Title: Identification and molecular characterization of missense mutations in orphan GPCR GPR61 occurring in severe obesity

doi: 10.1101/2024.10.21.24315834

Figure Lengend Snippet: PolyPhen-2 analysis of the 34 mutations in GPR61 to predict their impact on structure and function of the protein.

Article Snippet: HiBiT-GPR61 plasmid DNA was generated with Gibson cloning using GPR61-Tango plasmid DNA (#66366 Addgene, deposited by Bryan Roth) and HiBiT-FZD 6 plasmid DNA as templates.

Techniques:

Journal: medRxiv

Article Title: Identification and molecular characterization of missense mutations in orphan GPCR GPR61 occurring in severe obesity

doi: 10.1101/2024.10.21.24315834

Figure Lengend Snippet: NanoBiT-based measurement of cell surface expression of 34 mutants of GPR61. The red bars indicate statistically significant decrease and the green bars indicate statistically significant increase in NanoBiT-emmitted cell membrane-associated luminescence indicative of cell surface expression of HiBiT-tagged GPR61 constructs; at least p<0.05 and at least n=3 independent experiments.

Article Snippet: HiBiT-GPR61 plasmid DNA was generated with Gibson cloning using GPR61-Tango plasmid DNA (#66366 Addgene, deposited by Bryan Roth) and HiBiT-FZD 6 plasmid DNA as templates.

Techniques: Expressing, Membrane, Construct

Journal: medRxiv

Article Title: Identification and molecular characterization of missense mutations in orphan GPCR GPR61 occurring in severe obesity

doi: 10.1101/2024.10.21.24315834

Figure Lengend Snippet: A. EPAC-derived FRET-based biosensor reveals that three mutations (the red bars) lead to the statistically significant reduction in cAMP production elicited by overexpression of a GPR61 in the absence of an agonist (increase in the FRET ratio reflects decrease in cAMP levels). B. The presence of overexpressed GPR61 R236C leads to a statistically-significant reduction in the bystander BRET ratio between Gαs-67-RlucII and rGFP-CAAX indicative of reduced G s translocation away from the cell membrane; at least p<0.05 and at least n=3 independent experiments.

Article Snippet: HiBiT-GPR61 plasmid DNA was generated with Gibson cloning using GPR61-Tango plasmid DNA (#66366 Addgene, deposited by Bryan Roth) and HiBiT-FZD 6 plasmid DNA as templates.

Techniques: Derivative Assay, Over Expression, Translocation Assay, Membrane