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snap 94847  (MedChemExpress)
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MedChemExpress snap 94847
Co-expression and functional analysis of <t>MCHR1</t> and MRAP proteins in human and mouse RNA-seq datasets. (A, B) Co-expression correlation analysis of MRAP1 (A) or MRAP2 (B) with MCHR1 from the bulk RNA-seq data of human or mouse brain and hypothalamus. (C) Correlation coefficient between MRAP2 and MCHR1 from four single-cell RNA-seq datasets of mouse hypothalamus. (D) Changes in cell proportion of MCHR1 and MRAP2 in different metabolic states. One-way ANOVA with post-hoc Tukey test. **p < 0.01, ***p < 0.001. (E) GO enrichment pathway analysis of MCHR1 and MRAP proteins.
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Co-expression and functional analysis of MCHR1 and MRAP proteins in human and mouse RNA-seq datasets. (A, B) Co-expression correlation analysis of MRAP1 (A) or MRAP2 (B) with MCHR1 from the bulk RNA-seq data of human or mouse brain and hypothalamus. (C) Correlation coefficient between MRAP2 and MCHR1 from four single-cell RNA-seq datasets of mouse hypothalamus. (D) Changes in cell proportion of MCHR1 and MRAP2 in different metabolic states. One-way ANOVA with post-hoc Tukey test. **p < 0.01, ***p < 0.001. (E) GO enrichment pathway analysis of MCHR1 and MRAP proteins.

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Co-expression and functional analysis of MCHR1 and MRAP proteins in human and mouse RNA-seq datasets. (A, B) Co-expression correlation analysis of MRAP1 (A) or MRAP2 (B) with MCHR1 from the bulk RNA-seq data of human or mouse brain and hypothalamus. (C) Correlation coefficient between MRAP2 and MCHR1 from four single-cell RNA-seq datasets of mouse hypothalamus. (D) Changes in cell proportion of MCHR1 and MRAP2 in different metabolic states. One-way ANOVA with post-hoc Tukey test. **p < 0.01, ***p < 0.001. (E) GO enrichment pathway analysis of MCHR1 and MRAP proteins.

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Expressing, Functional Assay, RNA Sequencing Assay

Protein interaction of MCHR1 with MRAP2 but not MRAP1. (A) Tissue distribution of MCHR1, MRAP1, and MRAP2 tested by RT-PCR. β-actin was used as an internal control. (B) Expression abundance analysis of MCHR1, MRAP1, and MRAP2 in 14 mouse tissues. (C, D) Co-IP analysis of the interaction between 3HA-MCHR1 and 2Flag-MRAP1 (C) or 2Flag-MRAP2 (D) . The numbers on the right indicate molecular weight of marker band on the right (kd). (E, F) MCHR1-F1 co-localizes with MRAP1-Flag-F2 or MRAP2-Flag-F2 in live cells. YFP fluorescence is exhibited in green (left panel). MRAP1 or MRAP2 in the same cells detected with anti-Flag antibody and secondary anti-mouse Alexa594 is shown in red (middle panel). DAPI were applied to stain cell nuclei and shown in blue in merge figures (right panel) (scale bars, 10 μm). (G) Western blot for ERK1/2 and pERK1/2, and Tubulin is used as reference. The sample order: MCHR1, MCHR1+MRAP2, MCHR1 with MCH simulated, MCHR1+MRAP2 with MCH simulated (from left to right).

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Protein interaction of MCHR1 with MRAP2 but not MRAP1. (A) Tissue distribution of MCHR1, MRAP1, and MRAP2 tested by RT-PCR. β-actin was used as an internal control. (B) Expression abundance analysis of MCHR1, MRAP1, and MRAP2 in 14 mouse tissues. (C, D) Co-IP analysis of the interaction between 3HA-MCHR1 and 2Flag-MRAP1 (C) or 2Flag-MRAP2 (D) . The numbers on the right indicate molecular weight of marker band on the right (kd). (E, F) MCHR1-F1 co-localizes with MRAP1-Flag-F2 or MRAP2-Flag-F2 in live cells. YFP fluorescence is exhibited in green (left panel). MRAP1 or MRAP2 in the same cells detected with anti-Flag antibody and secondary anti-mouse Alexa594 is shown in red (middle panel). DAPI were applied to stain cell nuclei and shown in blue in merge figures (right panel) (scale bars, 10 μm). (G) Western blot for ERK1/2 and pERK1/2, and Tubulin is used as reference. The sample order: MCHR1, MCHR1+MRAP2, MCHR1 with MCH simulated, MCHR1+MRAP2 with MCH simulated (from left to right).

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Expressing, Co-Immunoprecipitation Assay, Molecular Weight, Marker, Fluorescence, Staining, Western Blot

Regulation of MCHR1 trafficking by WT MRAP2 and its mutants. (A) Surface expression of MCHR1 measured by ELISA assay, which transfected with MRAP2 at different ratios (from 1:0 to 1:9). (B) D-value of MCHR1 membrane surface expression in the control group (RAMP3) and MRAP2 group at different transfection ratios. (C) Total expression level of MCHR1. (D) Localization of GFP-MCHR1 by confocal microscopy, which transfected with empty vector (left) or MRAP2 (middle) or RAMP3 (right). (E) Sequence alignment of human and mouse MRAP2. (F) Schematic representation of the MRAP2 mutants constructed. (G) The surface expression of MCHR1 when co-transfected with empty vector (control), WT MRAP2, or different mutants. One-way ANOVA with post-hoc Tukey test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Regulation of MCHR1 trafficking by WT MRAP2 and its mutants. (A) Surface expression of MCHR1 measured by ELISA assay, which transfected with MRAP2 at different ratios (from 1:0 to 1:9). (B) D-value of MCHR1 membrane surface expression in the control group (RAMP3) and MRAP2 group at different transfection ratios. (C) Total expression level of MCHR1. (D) Localization of GFP-MCHR1 by confocal microscopy, which transfected with empty vector (left) or MRAP2 (middle) or RAMP3 (right). (E) Sequence alignment of human and mouse MRAP2. (F) Schematic representation of the MRAP2 mutants constructed. (G) The surface expression of MCHR1 when co-transfected with empty vector (control), WT MRAP2, or different mutants. One-way ANOVA with post-hoc Tukey test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Membrane, Control, Confocal Microscopy, Plasmid Preparation, Sequencing, Construct

Interaction between 3HA-MCHR1 and MRAP2 mutants. (A–D) The interaction between MCHR1 and MRAP2 C-terminal mutants. (E–H) The interaction between MCHR1 and MRAP2 N-terminal mutants. Middle lane with color is protein marker in all figures. The numbers on the right indicate molecular weight of marker band in the middle (kd).

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Interaction between 3HA-MCHR1 and MRAP2 mutants. (A–D) The interaction between MCHR1 and MRAP2 C-terminal mutants. (E–H) The interaction between MCHR1 and MRAP2 N-terminal mutants. Middle lane with color is protein marker in all figures. The numbers on the right indicate molecular weight of marker band in the middle (kd).

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Marker, Molecular Weight

Co-localization of MRAP2 mutants with MCHR1. (A) Confocal images of MCHR1 (green) when co-transfected with WTMRAP2 (red). (B–E) Localization of MCHR1 (green) when co-transfected with MRAP2 C-terminal mutants (red). (F–I) Images are co-transfected with MCHR1 (green) and MRAP2 N-terminal mutants. DAPI was used to stain cell nuclei and shown in blue. Merged images are shown in yellow.

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Co-localization of MRAP2 mutants with MCHR1. (A) Confocal images of MCHR1 (green) when co-transfected with WTMRAP2 (red). (B–E) Localization of MCHR1 (green) when co-transfected with MRAP2 C-terminal mutants (red). (F–I) Images are co-transfected with MCHR1 (green) and MRAP2 N-terminal mutants. DAPI was used to stain cell nuclei and shown in blue. Merged images are shown in yellow.

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Transfection, Staining

The influence of MCHR1 signaling by the functional domains of MRAP2. (A–H) Calcium response of MCHR1 simulated by increasing concentrations of MCH, transfected with WT MRAP2 or indicated mutants. (I–P) Competition binding assay of MCHR1 in 293T cells transfected with wt MRAP2 or indicated mutants. Relative luminescence intensity of NFAT-luc represents the normalized NFAT-luc units to p-RL-TK units (transfected internal control). Each data point represents the mean ± SEM of three replicates ( N = 3). (Q) Schematic diagram of distinct regions of MRAP2 required for regulating the trafficking and signaling of MCHR1.

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: The influence of MCHR1 signaling by the functional domains of MRAP2. (A–H) Calcium response of MCHR1 simulated by increasing concentrations of MCH, transfected with WT MRAP2 or indicated mutants. (I–P) Competition binding assay of MCHR1 in 293T cells transfected with wt MRAP2 or indicated mutants. Relative luminescence intensity of NFAT-luc represents the normalized NFAT-luc units to p-RL-TK units (transfected internal control). Each data point represents the mean ± SEM of three replicates ( N = 3). (Q) Schematic diagram of distinct regions of MRAP2 required for regulating the trafficking and signaling of MCHR1.

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Functional Assay, Transfection, Binding Assay, Control

Statistical analysis of MCHR1 in the presence of different MRAP2 mutants in response to MCH.

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Statistical analysis of MCHR1 in the presence of different MRAP2 mutants in response to MCH.

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Comparison

Statistical analysis of MCHR1 in the presence of different MRAP2 mutants in response to the antagonist of MCHR1.

Journal: Frontiers in Endocrinology

Article Title: Determination of the Interaction and Pharmacological Modulation of MCHR1 Signaling by the C-Terminus of MRAP2 Protein

doi: 10.3389/fendo.2022.848728

Figure Lengend Snippet: Statistical analysis of MCHR1 in the presence of different MRAP2 mutants in response to the antagonist of MCHR1.

Article Snippet: SNAP-94847 (MCHR1 antagonist) was purchased from MCE (MedChemExpress).

Techniques: Comparison