Journal: Biology of Sex Differences

Article Title: Sex-specific difference on anxiety- and depressive-like behavior in neuronal growth regulator 1-knockout mice

doi: 10.1186/s13293-025-00816-2

Figure Lengend Snippet: Negr1 –/– mice show sex-dependent characteristics. A. This schedule provides a schematic representation of the experimental design and indicates the behavioral test period (blue box). B. Negr1 protein expression in brain tissue, confirmed by Western blot. C. Body weight was measured over time, with values recorded on the day before each behavioral testing. D . Intestinal permeability assessed using plasma FITC–dextran levels. (E) Serum CORT levels were measured in Cohort 1 (at 8, 11, and 14 weeks of age). (F) Serum CORT levels were measured in Cohort 3 (at 32 weeks, endpoint). (G) Bdnf mRNA expression in the brain was measured in Cohort 2 (at 8 and 18 weeks of age). (H) Bdnf mRNA expression in the brain was measured in Cohort 3 (at 32 weeks, endpoint). RM-ANOVA was used for C , two-way ANOVA for D , F and H , and three-way ANOVA (age x sex x genotype) for E and G . Post hoc comparisons were performed using Sidak’s test. Male vs. female comparisons: ## p < 0.01 and ### p < 0.001. WT vs. Negr1 –/– comparisons: * p < 0.05, ** p < 0.01 and *** p < 0.01. Letters (a–c) denote the significant main effect of age. Data are presented as mean ± SEM. M, males; F, females

Article Snippet: The primary antibodies used included Negr1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), p-eIF2α (Cell signaling Technology, Danvers, MA, USA, 1:1000), eIF2α (Cell signaling Technology, 1:1000), caspase-3 (Cell signaling Technology, 1:1000), vinculin (Santa Cruz Biotechnology, 1:5000) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Thermo Fisher Scientific, 1:1000).

Techniques: Expressing, Western Blot, Permeability, Clinical Proteomics

Journal: Biology of Sex Differences

Article Title: Sex-specific difference on anxiety- and depressive-like behavior in neuronal growth regulator 1-knockout mice

doi: 10.1186/s13293-025-00816-2

Figure Lengend Snippet: Both sexes of Negr1 –/– mice exhibited anxiety, reduced sociability, and impaired spatial learning. (A) Assessment of center zone distance (%) and time (%) relative to total distance and time, measured in the open field test (OFT). (B) Assessment of social interaction ratio in the 3-chnaber social interaction test (3-SIT). (C) Assessment of total number of arm entries (left), the number of spontaneous alternations (middle), and the percentage of spontaneous alternations (right) in the Y-maze. (D) Latency to the hidden platform during the five-day Morris water maze (MWM) training phase (left), percentage of time in the target quadrant on day 6 (middle), and representative swim paths from the 18-week probe trial (right). Data in this figure were obtained from the Cohort 3 and summarize genotype effects across all tested ages (8–30 weeks). These assays showed a significant main effect of genotype but no significant sex x genotype interaction. The data are presented separated by sex to maintain consistency with the study’s focus on sex-specific differences. RM-ANOVA was used to assess group main effects and interactions, with Sidak’s post hoc comparisons. WT vs. Negr1 –/– comparisons: * p < 0.05, ** p < 0.01, and *** p < 0.001. Data are presented as mean ± SEM. M, males; F, females

Article Snippet: The primary antibodies used included Negr1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), p-eIF2α (Cell signaling Technology, Danvers, MA, USA, 1:1000), eIF2α (Cell signaling Technology, 1:1000), caspase-3 (Cell signaling Technology, 1:1000), vinculin (Santa Cruz Biotechnology, 1:5000) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Thermo Fisher Scientific, 1:1000).

Techniques:

Journal: Biology of Sex Differences

Article Title: Sex-specific difference on anxiety- and depressive-like behavior in neuronal growth regulator 1-knockout mice

doi: 10.1186/s13293-025-00816-2

Figure Lengend Snippet: Female Negr1 –/– mice exhibited greater depression-like behavior and impaired fear learning, whereas male Negr1 –/– mice exhibited greater anxiety. (A) Assessment of latency time in the passive avoidance test (PAT). (B) Assessment of immobility time in the tail suspension test (TST). (C) Assessment of open arm distance (%) and time (%) relative to total distance and time, measured in the elevated plus maze (EPM). Data in this figure were obtained from Cohort 3, which underwent repeated behavioral assessments (8–30 weeks). Panel A presents longitudinal data to reflect the significant age x sex x genotype interaction observed. Panel B and C present time-collapsed summary data across the tested ages, as only a significant sex x genotype interaction was observed without a significant age interaction. All data are presented separated by sex to highlight the significant sex x genotype interactions. RM-ANOVA was used to assess group main effects and interactions, with Sidak’s post hoc comparisons. WT vs. Negr1 –/– comparisons: ** p < 0.01. Data are presented as mean ± SEM. M, males; F, females

Article Snippet: The primary antibodies used included Negr1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), p-eIF2α (Cell signaling Technology, Danvers, MA, USA, 1:1000), eIF2α (Cell signaling Technology, 1:1000), caspase-3 (Cell signaling Technology, 1:1000), vinculin (Santa Cruz Biotechnology, 1:5000) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Thermo Fisher Scientific, 1:1000).

Techniques: Suspension

Journal: Biology of Sex Differences

Article Title: Sex-specific difference on anxiety- and depressive-like behavior in neuronal growth regulator 1-knockout mice

doi: 10.1186/s13293-025-00816-2

Figure Lengend Snippet: Negr1 –/– mice exhibit sex-specific regulation of ER stress. XBP1s mRNA expression in the brain, liver, and colon measured by RT-PCR. mRNA expression was quantified using ImageJ. Two-way ANOVA was used to assess group main effects and interactions, with Sidak’s post hoc comparisons. Male vs. female comparisons: ### p < 0.001. WT vs. Negr1 –/– comparisons: *** p < 0.001. Data are presented as mean ± SEM. M, males; F, females; †: non-specific band

Article Snippet: The primary antibodies used included Negr1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), p-eIF2α (Cell signaling Technology, Danvers, MA, USA, 1:1000), eIF2α (Cell signaling Technology, 1:1000), caspase-3 (Cell signaling Technology, 1:1000), vinculin (Santa Cruz Biotechnology, 1:5000) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Thermo Fisher Scientific, 1:1000).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Biology of Sex Differences

Article Title: Sex-specific difference on anxiety- and depressive-like behavior in neuronal growth regulator 1-knockout mice

doi: 10.1186/s13293-025-00816-2

Figure Lengend Snippet: Negr1 –/– mice exhibit sex-specific regulation of apoptosis. Caspase-3 protein expression in the brain, liver, and colon measured by Western blot and quantification of caspase-3 protein levels in tissues. Normalization to loading controls for brain pro-caspase-3 (left) and cleaved/pro-caspase-3 ratios in liver (middle) and colon (right). Protein expression were quantified in ImageJ. Two-way ANOVA was used to assess group main effects and interactions, with Sidak’s post hoc comparisons. Male vs. female comparisons: # p < 0.05 and ## p < 0.01. WT vs. Negr1 –/– comparisons : * p < 0.05 and *** p < 0.001. Data are presented as mean ± SEM. M, males; F, females

Article Snippet: The primary antibodies used included Negr1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:1000), p-eIF2α (Cell signaling Technology, Danvers, MA, USA, 1:1000), eIF2α (Cell signaling Technology, 1:1000), caspase-3 (Cell signaling Technology, 1:1000), vinculin (Santa Cruz Biotechnology, 1:5000) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Thermo Fisher Scientific, 1:1000).

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A) NEGR1+/+ cultured hypothalamic neuron immunolabelled for NEGR1, GAD65 and VGAT. Magnified images of the outlined areas show examples of NEGR1 clusters overlapping with GAD65 and VGAT accumulations (arrows). NEGR1-/- neurons were labelled for control. Bars, 10 µm. (B) NEGR1+/+ cultured hypothalamic neurons immunolabelled for a dendritic marker protein MAP2 and axonal marker protein tau. Bar, 20 µm. (C-F) Western blot (WB) analysis of VGAT ( C, D ) and GAD65 ( E, F ) in NEGR1+/+ and NEGR1-/- brain homogenates (H), synaptosomes (S) and soluble protein (sol) fractions. NEGR1 labelling was included in ( C ) to illustrate its enrichment in synaptosomes. Two different exposures of GAD65 labelling in ( E ) are shown to better illustrate a reduction in GAD65 levels in NEGR1-/- synaptosomes and soluble protein fractions. Labelling for GAPDH and Ponceau stain was used to control loading. Labelling for synaptophysin was used to confirm the efficiency of synaptosome isolation. Graphs show mean ± SEM changes in protein enrichment in NEGR1-/- synaptosomes and soluble protein fractions vs enrichment in NEGR1+/+ samples set to 1. n = 5 pairs of NEGR1+/+ and NEGR1-/- mice were analyzed. *p, one sample t test vs +/+. ( G ) NEGR1+/+ and NEGR1-/- cultured hypothalamic neurons immunolabeled for GAD65 and synaptophysin. ( H ) NEGR1-/- cultured hypothalamic neurons transfected with GAD65-GFP and pcDNA3 or NEGR1 immunolabelled for synaptophysin and NEGR1. In G , H , examples of synaptophysin positive (arrowheads) and negative (arrows) GAD65 clusters are shown. Bars, 5 µm. Graphs show cumulative frequency (CF) of neurons with different percentages of synaptophysin negative GAD65 clusters. *p, Mann-Whitney test (n = 20 ( G ) and 30 ( H ) neurons per group). Figure 1 source data. Contains uncropped Western blot source data for Figure 1.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Cell Culture, Control, Marker, Western Blot, Staining, Isolation, Protein Enrichment, Immunolabeling, Transfection, MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A) NEGR1+/+ cultured hypothalamic neurons treated with recombinant soluble NEGR1 (sNEGR1) or mock-treated with the culture medium (control) immunolabeled for GAD65 and synaptophysin. Bar, 5 µm. Graph shows mean + SEM levels of GAD65 in synaptophysin accumulations relative to the mean value of the control set to 100% (n > 1000 boutons analyzed per group). *p, one-way ANOVA and Dunnett’s multiple comparisons test (compared to control). (B) Western blot (WB) analysis of GAD65 levels in homogenates (H), synaptosomes (S) and soluble protein fractions (sol) from hypothalamic tissues treated with BSA (control) or sNEGR1. GAPDH and synaptophysin served as loading controls. Graphs show mean ± SEM GAD65 levels in sNEGR1-treated tissues relative to GAD65 levels in the control set to 100% from n = 3 independent experiments. *p, one sample t test vs control. (C) Concentration of GABA in synaptosomes from hypothalamic tissues treated with BSA (control) or sNEGR1 (n = 7). *p, t test. Figure 2 source data. Contains uncropped Western blot source data for Figure 2.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Cell Culture, Recombinant, Control, Immunolabeling, Western Blot, Concentration Assay

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A-D) Synaptic vesicles in cultured hypothalamic neurons were loaded with FM4-64 applied for 2 min in 47 mM K+ buffer (0 s). Time lapse images show release of the dye in response to the electric field stimulation. Bar, 5 μm. Graphs show FM4-64 uptake at 0 s, changes in FM4-64 levels (ΔF/F) in synaptic boutons over time during the stimulation, and half-life of FM4-64 loss during the stimulation (mean ± SEM, *p, Mann-Whitney test). (A) NEGR1+/+ neurons (n = 113 boutons from 10 neurons) and NEGR1-/- neurons (n = 108 boutons from 10 neurons). (B, C) NEGR1+/+ neurons co-transfected with GFP and pcDNA3 or NEGR1. Axons ( B, n = 418 boutons from 10 pcDNA3-transfected neurons, 266 boutons from 12 NEGR1-transfected neurons) and dendrites ( C , n = 376 boutons from 12 pcDNA3-transfected neurons, 261 boutons from 13 NEGR1- transfected neurons). (D) NEGR1+/+ neurons treated with control non-specific immunoglobulins (Ig, n = 681 boutons from 10 neurons) or NEGR1 antibodies (NEGR1 Ab, n = 514 boutons from 10 neurons).

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Cell Culture, MANN-WHITNEY, Transfection, Control

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A, B) NEGR1+/+ cultured hypothalamic neurons treated with sNEGR1 and dynasore ( A ) or TetTx ( B ) as indicated. Control neurons were mock treated with vehicle (DMSO in A , water in B ). Neurons were immunolabeled for GAD65 and synaptophysin. Examples of synaptophysin positive (arrows) and negative (arrowheads) GAD65 clusters are shown. Bars, 5 µm. Graphs show cumulative frequency (CF) of neurons with increasing percentages of synaptophysin negative GAD65 clusters. *p, Mann-Whitney test (n = 20-26 neurons per group). (C, D) sNEGR1-treated and control culture medium-treated NEGR1+/+ cultured hypothalamic neurons incubated live with antibodies against the lumenal domain of VGAT. The antibodies were detected with secondary antibodies applied before (VGAT surface_remaining ) and after (VGAT surface_delivered ) permeabilization of membranes with detergent. Neurons were co-labelled with antibodies against the cytoplasmic domain of VGAT (VGAT synaptic ). Bar, 5 μm. Graphs show mean + SEM labelling intensities of VGAT synaptic and VGAT surface_delivered ( C ), VGAT surface_remaining ( D ), and their ratios (n = 40 neurons in C , 20 neurons in D ). Control mean intensity was set to 100%. (E) Western blot analysis of GAD65 levels in synaptic PM from hypothalamic tissues treated with BSA or sNEGR1 applied either alone (cont.) or together with dynasore (dyn.) or TetTx. NCAM1 served as loading control. Graph shows mean ± SEM GAD65 levels relative to the levels in BSA only treated tissues set to 100% from n = 3 independent experiments. Samples from each experiment were analyzed by Western blot twice and the data were averaged. *p, one sample t test vs BSA only treated tissues. Figure 4 source data. Contains uncropped Western blot source data for Figure 4.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Cell Culture, Control, Immunolabeling, MANN-WHITNEY, Incubation, Western Blot

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A) Diagram illustrating the principle of detection of the PM attachment of GAD65 using BiFC. (B) LCK-VN-transfected CHO cell labelled with VN-recognizing GFP antibodies and co-labelled for GM130. (C) BiFC assay with CHO cells co-transfected with LCK-VN and GAD65-VC proteins shown on the diagram. Immunolabelling for GAD65 was used for normalization. Graph shows mean ± SEM BiFC / GAD65 ratio. *p, one-way ANOVA and Dunnett’s multiple comparisons test (n = 15 cells per group). (D) BiFC fluorescence distribution in the XY confocal slice and ZX and ZY sections along the dashed lines through the 3D reconstructed GAD65WT-VC-co-transfected cell shown in C . Note PM localization of the BiFC signals in ZX and ZY sections. Arrowheads show vesicle-like structures at the PM. (E) CHO cells co-transfected with LCK-VN, GAD65WT-VC and either pcDNA3 or NEGR1. Cells were co-labelled for NEGR1. Arrows show clusters of NEGR1 co-localized with BiFC signals. (F) CHO cell co-transfected with LCK-VN and GAD65WT-VC co-labelled for clathrin. Arrowheads in ZX and ZY sections through the 3D reconstructed cell show BiFC signals co-localized with clathrin accumulations at the PM. (G) FM4-64 loaded vesicles in CHO cells co-transfected with GAD65-GFP and either pcDNA3 or NEGR1. High magnification images show areas outlined with dashed boxes. Arrows show examples of FM-loaded vesicles co-localized with GAD65-GFP. Graph shows mean ± SEM percentages of FM- loaded vesicles co-localized with GAD65-GFP and FM4-64 levels. *p, Mann-Whitney test (n = 42 cells per group). Bars, 20 µm (low magnification), 5 µm (high magnification) (B-G) .

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Transfection, Bimolecular Fluorescence Complementation Assay, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A, B) CHO cells transfected with pcDNA3 or NEGR1 were either co-transfected with mCherry-D4H to visualize cholesterol (A) or immunolabelled for ganglioside GM4 (B) . Bars, 20 µm. Graphs show ratios of D4H and GM4 labelling levels at the PM and in cytoplasm (mean + SEM, n = 20). *p, Mann Whitney test. (C) Lipid raft markers gangliosides GM4 and GD3 and phosphatidylinositol 4,5-bisphosphate (PIP2) analyzed in triplicate by dot blot in brain homogenates (BH) and synaptosomes (synapt.) from a representative pair of NEGR1+/+ and NEGR1-/- littermates. Graphs show mean + SEM synaptic enrichments of the lipids in NEGR1-/- mice relative to the levels in NEGR1+/+ mice set to 1. n = 5 pairs of NEGR1+/+ and NEGR1-/- mice were analyzed. *p, one sample t test vs +/+. Figure 6 source data. Contains dot blot source data for Figure 6.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Transfection, MANN-WHITNEY, Dot Blot

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: ( A ) Neurites of NEGR1+/+ cultured hypothalamic neurons immunolabelled for NEGR1, GAD65 and NPY. Arrows show examples of NEGR1 clusters overlapping with NPY positive GAD65 accumulations. Open arrowheads show NPY negative GAD65 accumulations with lower levels of NEGR1. Bar, 5 µm. Graph shows mean ± SEM levels of NEGR1 in NPY positive (+) and negative (-) GAD65 accumulations (n > 100). AU – arbitrary units defined as pixel values of 16-bit gray scale images. *p, unpaired t test. ( B, C ) Confocal images of the ARC ( B ) and pyramidal cell layer in the hippocampal CA3 region ( C ) in brain sections of NEGR1+/+ and NEGR1-/- littermates immunolabelled for NEGR1, GAD65 and VGAT. Bar, 10 µm. Graphs show mean ± SEM densities of VGAT accumulations. *p, Mann-Whitney test.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Cell Culture, MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: ( A ) Mean ± SEM percentages of pellets eaten over 30 min during magazine training sessions. One pellet was released into the magazine each minute. ( B ) Mean ± SEM numbers of active lever presses and pellets eaten during the fixed ratio task. Each active lever press led to the release of a pellet. ( C ) Individual progressive ratio break point values measured over three consecutive days in the progressive ratio task. Lines connect data for individual mice. Graphs showing data normalized to the values on day 1 set to 100% are included to illustrate an increase in the progressive ratio break point in NEGR1+/+ mice. ( D ) Mean ± SEM of the normalized values shown in C . ( E ) Mean ± SEM numbers of pellets delivered and active lever presses in the progressive ratio task. The numbers of active lever presses were normalized to the number on day 1 set to 100%. In A - E , n = 9 NEGR1+/+, 8 NEGR1+/- and 12 NEGR1-/- mice were analyzed. *p, repeated measures two-way ANOVA and Tukey’s multiple comparisons test.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques:

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A-C) Western blot (WB) analysis of NEGR1 levels in homogenates (H), synaptosomes (S) and soluble protein fractions (sol) from brains of chow and high fat diet (HFD) fed mice. GAPDH served as loading control. Graphs show levels of full length NEGR1 in brain homogenates and synaptosomes ( B ) and soluble NEGR1 fragments in the soluble protein fraction ( C ) from HFD fed mice relative to the levels in chow diet fed mice set to 100%. n = 3 mice per group. ( D ) WB analysis of the soluble protein fractions from NEGR1+/+ and NEGR1-/- brains with NEGR1 antibodies. Note that the NEGR1 fragment is detected only in the NEGR1+/+ lane. The diagram shows the structure of NEGR1 consisting of three Ig-domains attached to the PM via a GPI anchor. The site recognized by the antibody used for Western blot analysis is marked with a square. ( E ) WB analysis of GAD65 levels in synaptic PM from chow and HFD fed mice. NCAM1 served as loading control. Graph shows levels in HFD membranes relative to chow control set to 100%. n = 3 mice per group. (F) Concentration of GABA in synaptosomes from the brains of chow and HFD fed mice (n = 10). (G) WB analysis of GAD65 levels in homogenates (H), synaptosomes (S) and soluble protein fractions (sol) from brains of chow and HFD fed mice. GAPDH and synaptophysin served as loading controls. Graphs show levels of GAD65 in brain homogenates and synaptosomes of HFD fed mice relative to the levels in chow fed mice set to 100%. n = 3 mice per group. (H) Axons and dendrites of NEGR1+/+ cultured hypothalamic neurons co-transfected with cherry and control pcDNA3 vector or NEGR1. Neurons were immunolabelled for synaptophysin and NEGR1. Note reduced numbers of synaptophysin puncta (arrows) along the axons and dendrites of NEGR1-overexpressing neurons. Bar, 5 µm. Graphs show synaptophysin labelling intensities along dendrites (n = 14 - 24) and axons (n = 31 - 43). Mean of control was set to 100%. In B, C, E-H , mean ± SEM values are shown. *p, one sample t test vs chow ( B, C, E, G ) or Mann Whitney test ( F, H ). Figure 10 source data. Contains uncropped Western blot source data for Figure 10.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Western Blot, Control, Concentration Assay, Cell Culture, Transfection, Plasmid Preparation, MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuronal growth regulator 1 (NEGR1) promotes synaptic targeting of glutamic acid decarboxylase 65 (GAD65)

doi: 10.1101/2022.02.08.479601

Figure Lengend Snippet: (A) Synaptic vesicle localized GAD65 uses glutamate to synthesize GABA, which is transported to the vesicle lumen by VGAT ( a ). GAD65 is de-palmitoylated and removed to the cytosol from refilled vesicles ( b ), which then fuse with the PM and release GABA. GAD65 released to the cytosol can travel to the Golgi in the neuronal cell body (red arrow, ) or attach to the PM in the synapse ( c ). Palmitoylation of GAD65 at the PM targets it to the NEGR1-containing cholesterol-enriched microdomains, which are anchored in synapses by NEGR1-containing adhesive bonds ( d ). NEGR1-dependent synaptic clustering of lipid rafts promotes ‘loading’ of palmitoylated GAD65 on the newly formed synaptic vesicles ( e ). This loading is facilitated by interactions between NEGR1 containing lipid rafts and components of synaptic vesicles constraining the retrieval of synaptic vesicle membranes. Synaptic recycling of vesicles and GAD65 is shown with black and pink arrows, respectively. (B) In NEGR1+/+ neurons, synaptic clustering of lipid rafts by trans-synaptic NEGR1-containing adhesive bonds promotes synaptic accumulation of GAD65. (C) In NEGR1-/- neurons, synaptic clustering of lipid rafts is reduced, synaptic targeting of GAD65 is inhibited, and non-synaptic GAD65 clusters are formed.

Article Snippet: Goat polyclonal antibodies against NEGR1 (AF5394) used for immunocytochemistry (IC: 1:100) and immunohistochemistry (IHC: 1:1000) were from R&D systems.

Techniques: Adhesive

Journal: Frontiers in Immunology

Article Title: Patients’ IgLON5 autoantibodies interfere with IgLON5-protein interactions

doi: 10.3389/fimmu.2023.1151574

Figure Lengend Snippet: Membrane proteins identified as bona-fide IgLON5 interactors.

Article Snippet: The presence of the interacting proteins was detected by incubating the membrane with the following antibodies (dilution 1:500, overnight at 4°C): IgLON1 (STJ94590), IgLON2 (STJ94442), IgLON3 (STJ116461), IgLON4 (STJ94394, St. John’s Laboratory, London, UK), KIDINS220 (21856-1-AP, Proteintech, Rosemont, IL, USA), IGSF21 (21465-1-AP; Proteintech), CACNA2D2 (SAB1401461, SIGMA) and CASPR2 (ab33994, Abcam) followed by incubation with appropriate secondary antibodies (dilution 1:1000) for 1 hour at room temperature.

Techniques: Membrane

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 1 The IL-6 mRNA levels were increased in the eWAT of Negr1−/−mice (A) Quantitative RT-PCR was performed to measure the mRNA expression levels of indicated genes using total RNA isolated from the epididymal WAT (eWAT) of 11-week-old WT and Negr1−/−C57BL6 mice (n = 8). The data represent mean ± SEM. **p < 0.01, ***p < 0.001. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. (B) Quantitative RT-PCR analyses using total RNA obtained from bone-marrow-derived macrophages (n = 4). *p < 0.05. (C) Immunostaining of macrophages in eWAT. The paraffin-embedded WAT tissue sections were incubated with anti-F4/80 (red) and anti-CD80 (green) antibodies. Imaging was performed using an Olympus BX51 microscope. Fluorescence intensities were determined using ImageJ software. ***p < 0.001. (D, E) After blood samples were collected from 12-week-old mice (n = 8), the levels of IL-6 (D) and sIL-6R(E) were determined using the ELISA kit. The data represent mean ± SEM. *p < 0.05.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Quantitative RT-PCR, Expressing, Isolation, Derivative Assay, Immunostaining, Incubation, Imaging, Microscopy, Fluorescence, Software, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 2 The protein expression levels of IL-6R components in the eWAT of Negr1−/−mice. (A) Comparison of protein levels of IL-6 signaling complex in the gonadal WAT of WT and Negr1−/−mice (n = 9). The band intensity of each protein was normalized to the vinculin level. The data represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. (B) Visualization of IL-6R in the eWAT of 13-week-old WT and Negr1−/−mice by immunostaining paraffin-embedded tissue sections using an anti-IL-6R antibody (×400 magnification). Fluorescence signals were measured using ImageJ software.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Expressing, Comparison, Immunostaining, Fluorescence, Software

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 3 Increased STAT3 activation in the eWAT of Negr1−/−mice. (A) Immunostaining of eWAT tissue sections with an anti-pSTAT3 antibody. Imaging was performed using an Olympus BX51 microscope. Fluorescence intensity was calculated using ImageJ software. *p < 0.05. (B) Primary adipocytes (n = 4) were isolated from WT and Negr1−/−mice and incubated with IL-6 (25 ng/mL) for 10 min before immunoblotting. (C) Primary adipocytes were incubated with hyper-IL-6-containing conditioned media for 10 min and pSTAT3/STAT3 levels were calculated based on the band intensities using ImageJ software. **p < 0.01.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Activation Assay, Immunostaining, Imaging, Microscopy, Fluorescence, Software, Isolation, Incubation, Western Blot

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 4 NEGR1 may interact with IL-6R. (A) Protein structure of NEGR1 and IL-6R. (B) Co-immunoprecipitation (IP) was performed using an anti-FLAG antibody after HeLa cells were co-transfected with GFP-NEGR1 and FLAG-IL-6R for 24h. Co-isolated NEGR1 was visualized with the anti-GFP antibody. (C) Reciprocal co-IP between GFP-NEGR1 and FLAG-IL-6R using anti-GFP antibody. (D) Co-IP between the endogenous proteins. After HeLa cell lysates were subjected to IP with an anti-NEGR1 antibody, IL-6R was visualized with IL-6R antibody. (E, F) In situ proximity ligation assay of SKOV-3-FLAG- NEGR1 stable cells under permeabilized (E) or non-permeabilized (F) conditions. For cell permeabilization, cells were treated with 0.1% Triton X-100 for 10 min. Then, cells were incubated with rabbit anti-FLAG antibody and mouse anti-IL-6R antibody for 1 h, followed by treatment of PLA probes (anti- mouse MINUS and anti-rabbit PLUS). The bar represents 50 μm.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Immunoprecipitation, Transfection, Isolation, Co-Immunoprecipitation Assay, In Situ, Proximity Ligation Assay, Incubation

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 5 Domain mapping of the interaction between NEGR1 and IL-6R. (A) Domain structure of NEGR1 and IL-6R. (B) Interaction between GST-fused IL-6R extracellular domain (ECD, 20–357) and FLAG-tagged soluble NEGR1 (34–323) was examined by GST-pulldown. (C) Various IL-6R deletion constructs were generated and GST-pulldown was performed after 293T cells were transfected with IL-6R deletion mutants together with FLAG-NEGR1 (34–323). CBM, cytokine binding module; D1 ~ D3, domain 1–3; TM, transmembrane domain; CD, cytoplasmic domain. (D) Determination of NEGR1 domain required for IL-6R interaction. GST-pulldown was carried out after 293T cells were co-transfected with NEGR1 domain mutants and FLAG-IL-6R (20–357). C1 ~ C3, C2-type immunoglobulin domain 1–3.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Construct, Generated, Transfection, Binding Assay

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 6 Co-localization analysis of NEGR1 and IL-6R. (A) Gel filtration chromatography was performed with the Sephacryl S-400 HR column using SKOV-3- NEGR1-FLAG cells. Eluents were used for immunoblotting with anti-FLAG and anti-IL-6R antibodies. (B) Lipid raft fractionation using OptiPrep™ Gradient. Centrifugation was performed at 76,000 × g for 18 h at 4°C after 293T cells were transfected with FLAG-IL-6R together with GFP-NEGR1 (right) or GFP control (left). Frotillin-1 was used as a lipid raft marker. (C) Co-localization of NEGR1 and IL-6R in 3T3-L1-FLAG-NEGR1 stable cells. After cell permeabilization, cells were incubated with anti-FLAG (blue) and anti-IL-6R (red) antibodies. Cholera toxin subunit B (CTB) was used to visualize lipid rafts (green). Imaging was performed using a confocal laser scanning microscope Zeiss LSM 880 (Zeiss, Germany). The scale bar represents 50 μm. (D) Confocal microscopy of N2a cells after transfection of FLAG-NEGR1 plasmids. Cells were incubated using anti-FLAG (green) and anti-IL-6R (red) antibodies, and cell nuclei were stained with DAPI. The scale bar represents 50 μm.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Chromatography, Western Blot, Fractionation, Gradient Centrifugation, Transfection, Control, Marker, Incubation, Imaging, Laser-Scanning Microscopy, Confocal Microscopy, Staining

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 7 NEGR1 may attenuate IL-6 trans-signaling. (A) Hyper-IL-6-containing conditioned medium was obtained after transfection of HeLa cells with pcDNA3-hyper-IL-6-MYC for 24h. (B) After transfection with GFP-NEGR1, HeLa cells were incubated with Hyper-IL-6-containing medium for 10 min. Then, cells were lysed and used for immunoblotting with indicated antibodies. Band density was calculated using ImageJ software. (C) After HeLa cells were transfected with increasing amounts of GFP-NEGR1 plasmids, the cell lysates were mixed with the hyper-IL-6-containing conditioned medium. Then, IP was performed using an anti-gp130 antibody, and the co-isolated hyper-IL-6 was detected with an anti-MYC antibody. (D) Comparison of binding of Hyper-IL-6-MYC on the cell surface. After SKOV-3 (upper panels) and SKOV-3-FLAG-NEGR1 stable cells (lower panels) were incubated with hyper-IL-6 containing conditional medium 1h, cell surface-bound hyper-IL-6 protein was visualized using anti-MYC. Imaging was performed using an Olympus BX51 microscope (Tokyo, Japan). Integrated fluorescence intensity was quantified using ImageJ software. *p < 0.05. Scale bar = 50 mm.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques: Transfection, Incubation, Western Blot, Software, Isolation, Comparison, Binding Assay, Imaging, Microscopy

Journal: Frontiers in molecular biosciences

Article Title: Neuronal growth regulator 1 may modulate interleukin-6 signaling in adipocytes.

doi: 10.3389/fmolb.2023.1148521

Figure Lengend Snippet: FIGURE 8 Proposed role of NEGR1 in IL-6 signaling.

Article Snippet: The following antibodies were used to visualize specific proteins: FLAG, MYC, and β-actin (Sigma-Aldrich, St. Louis, MO, United States); GFP, IL-6R, gp130, NEGR1, AKT1, and p-AKT1 (Santa Cruz Biotechnology, Santa Cruz, CA, United States); glyceraldehyde 3- phosphate dehydrogenase (GAPDH; Cusabio, College Park, MD, USA); STAT3, p-STAT3 (Cell Signaling Technologies, Beverley, MA, United States), and human Fc (hFc; Gibco, Thermo Fisher Scientific, Waltham, MA, United States).

Techniques:

Journal: Cardiovascular Diabetology

Article Title: Expression of fourteen novel obesity-related genes in zucker diabetic fatty rats

doi: 10.1186/1475-2840-11-48

Figure Lengend Snippet: Primer and probes

Article Snippet: NEGR1 , Neuronal growth regulator 1 , Rn00572380_m1.

Techniques: Derivative Assay, Variant Assay

Journal: Cardiovascular Diabetology

Article Title: Expression of fourteen novel obesity-related genes in zucker diabetic fatty rats

doi: 10.1186/1475-2840-11-48

Figure Lengend Snippet: Expression of genes with a known neuronal expression. a) Relative expression of TMEM18. * p < 0.05 vs. ZL KF, ZL SF and ZL MF; † p < 0.05 vs. ZDF KF, ZDF SF and ZDF MF. b) Relative expression of KCTD15. * p < 0.05 vs. ZL KF and ZL SF; † p < 0.05 vs. ZDF KF and ZDF SF. c) Relative expression of NEGR1. * p < 0.05 vs. ZL KF, ZL SF and ZL MF; † p < 0.05 vs. ZDF KF, ZDF SF and ZDF MF; # p < 0.05 vs. ZDF KF; ¶ p < 0.05 vs. ZDF SF; ‡ p < 0.05 vs. ZDF MF. d) Relative expression of NRXN3. No detectable mRNA-amount in adipose tissues.

Article Snippet: NEGR1 , Neuronal growth regulator 1 , Rn00572380_m1.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: Differentially expressed genes and Kyoto Encyclopedia of Genes and Genomes analysis. Volcano plots showing the top twenty up- or downregulated genes for each cell type. The red dots represent upregulated genes, and the blue dots represent downregulated genes. ( P value < 0.05, and |log2foldchange| > 0.58). Bubble plot indicating the top enriched pathways for each cell type based on KEGGpathway enrichment analysis of differentially expressed genes( www.kegg.jp/kegg/kegg1.html ). The sizes of the dots represent the number of genes included in each pathway. The colour gradient of dots represents the adjusted P values of each enriched pathway. The genes Pfkfb3 , Tbc1d1 and the insulin signaling pathway; Pdgfd and the PI3K-Akt signaling pathway; Cxcr2 and PLD, Rap1 signaling pathways; Ccl21 and chemical carcinogenesis-reactive oxygen species signaling pathway; Mef2c and the calcium signaling pathway; Negr1 and leukocyte transendothelial migration signaling pathway are labelled by the black and red bars in Panels a , b , c , d , e , and f .

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques: Protein-Protein interactions, Migration

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: Protein-protein interaction network (PPI network).We constructed nine PPI networks displaying protein-protein interactions among the related genes. The nodes represent proteins, and the edges represent the interaction strength between two proteins. The proteins PFKFB3 ( a ), PDGFD ( b ), CXCR2 ( c ), CCL21 ( d ), SEMA3A ( e ), RYR3 ( f ), MEF2C ( g ), NEGR1( h ) and TBC1D1 ( i ) which are located at the hub of the interaction network, are responsible for diaphragm fibrosis and atrophy.

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques: Construct, Protein-Protein interactions

Journal: Scientific Reports

Article Title: Single-nucleus transcriptomic profiling of the diaphragm during mechanical ventilation

doi: 10.1038/s41598-024-82530-4

Figure Lengend Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. qRT-PCR and Western blotting showing the expression of selected genes and corresponding proteins in the mechanically ventilated diaphragm and control ones. The bar graphs show the quantification of the mRNA expression of Pfkfb3 ( a ), Pdgfd ( b ), Cxcr2 ( c ), Negr1 ( d ), Sema3a ( e ), and Mef2c ( f ) normalized to that of GAPDH. Asterisks indicate significant differences ( n = 3 in each group) (* P < 0.05). Western blotting analysis of the protein expression of PFKFB3 ( g ), PDGFD ( h ), CXCR2 ( i ), NEGR1 ( j ), SEMA3A ( k ), and MEF2C ( l ) normalized to that of β-actin as a loading control.

Article Snippet: After the semidry blotting procedure (50 min, 90 V), the membrane was incubated for 1 h at room temperature (RT) in 5% BSA blocking solution, followed by overnight incubation on a shaker at 4 °C with primary antibodies against PFKFB3 (bs-3528R, Boster Biological Technology Co., Ltd., Wuhan, China), PDGFD (bs-24572R, Boster), CXCR2 (bs-1629R, Boster), NEGR1 (bs-11095R, Boster), SEMA3A (bs-10468R, Boster), MEF2C (bs-4130R, Boster) and β-actin (66009-1-Ig, Proteintech, Wuhan, China) as a loading control.

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Expressing, Control

Journal: BioMed Research International

Article Title: Copy Number Variations in Candidate Genes and Intergenic Regions Affect Body Mass Index and Abdominal Obesity in Mexican Children

doi: 10.1155/2017/2432957

Figure Lengend Snippet: Copy number variants selected in five genes and four intergenic regions.

Article Snippet: Hs06530590_cn , esv3586400 , Chr1: 72516113–72546537 , NEGR1 , Intron 1 , [ ] .

Techniques:

Journal: BioMed Research International

Article Title: Copy Number Variations in Candidate Genes and Intergenic Regions Affect Body Mass Index and Abdominal Obesity in Mexican Children

doi: 10.1155/2017/2432957

Figure Lengend Snippet: Effect of copy number in genes and intergenic regions on body mass index and waist circumference.

Article Snippet: Hs06530590_cn , esv3586400 , Chr1: 72516113–72546537 , NEGR1 , Intron 1 , [ ] .

Techniques:

Journal: BioMed Research International

Article Title: Copy Number Variations in Candidate Genes and Intergenic Regions Affect Body Mass Index and Abdominal Obesity in Mexican Children

doi: 10.1155/2017/2432957

Figure Lengend Snippet: Association between the CNVs and abdominal obesity in Mexican children. The forest plot shows the results of the logistic regression models adjusted for age and gender; for the ARHGEF4 , NEGR1, and CPXCR1 and intergenic regions the reference group was defined as individuals with loss copy number (0–2) and compared with the group with the highest copy number (≥3), while for the INS the reference group was of children with the highest copy number (≥2) and compared with the group with loss copy number (0-1).

Article Snippet: Hs06530590_cn , esv3586400 , Chr1: 72516113–72546537 , NEGR1 , Intron 1 , [ ] .

Techniques: