nrcam concentrations Search Results


95
Thermo Fisher gene exp nrcam mm00663607 m1
Bar graphs and receiver operator curves of Neuronal Cell Adhesion Molecule <t>(NrCAM)</t> in two large cohorts ( a-d , Cohort 1, e-h , Cohort 2). NrCAM concentrations significantly reduced ( a , p = 4.75 x 10 −6 ) in the circulation of women at 36 weeks' gestation who subsequently delivered an FGR infant (birthweight <3rd centile). The discriminatory power of NrCAM is shown as a receiver operating characteristic (AUROC) curve with an area of the curve of 0.76 ( b ). Plasma Placental Growth Factor ( c, d ; PlGF) concentrations at 36 weeks were also significantly reduced in the same cohort ( c , p = 8.15 × 10 −7 ). The AUROC of PlGF is 0.77 ( d ). Cohort, n = 957 controls, n = 26 FGR. In a high-risk international cohort (FEMINA, Manchester, UK) ( e , f ) show NrCAM concentrations reduced ( e , p = 9.34 x 10 −3 ) in the circulation of women who presented at the clinic with reduced fetal movements who subsequently delivered an FGR infant (birthweight <3rd centile), AUROC of 0.72 ( f ). Plasma PlGF ( g, h ) concentrations in the same cohort remain unchanged ( g , p = 0.13). The AUROC of PlGF is 0.63 ( h ). Cohort, n = 235 controls, n = 12 FGR. groups compared using Mann–Whitney U tests. AUROC, area under the AUROC curve, with 95% confidence intervals presented in brackets. Data depicted for a , c, e and g are mean ± SEM. ∗∗p < 0.01, ∗∗∗∗p < 0.0001.
Gene Exp Nrcam Mm00663607 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nrcam+concentrations/Gene+Exp%2E+Nrcam%2C+Mm00663607_m1/pmc12301788-106-29-5
Average 95 stars, based on 1 article reviews
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93
R&D Systems mouse recombinant nrcam
Fig. 3. Expression and cleavage of selected gelatinase substrates in TNF- α and unstimulated WT astrocyte cultures. (A) Immunofluorescence staining for MMP substrates, VCAM-1, <t>NrCAM,</t> agrin, NOTCH3, together with GFAP to mark astrocytes and DAPI; boxed areas are shown to the right at higher magnifications. Scale bars, 100 μm. (B) Silver-stained gels showing cleavage products of gelatinase substrates after overnight incubation without (0) or with 1:10 or 1:100 ratios of MMP-9:substrate or ADAM10:substrate. Arrows mark the positions of ADAM10 in samples. Asterisks mark specific cleavage products. Data are representative of two to three experiments.
Mouse Recombinant Nrcam, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nrcam+concentrations/Recombinant+Mouse+NrCAM+Protein%2C+CF/pm37467333-294-44-47
Average 93 stars, based on 1 article reviews
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91
R&D Systems polyclonal rabbit antibodies against nrcam
Fig. 1 Immunolabeling of <t>NrCAM</t> in the mouse cochlea at E14 and E17. Cell nuclei are in blue, NrCAM in red, Hes5 in green. a, c Overview of sagittal section of mouse head at the embryonic stage E14 and E17, stained with Hematoxylin/Eosine. Boxed is the cochlea. b The diagram of the embryonic OC. Source: http://syllabus.med.unc. edu. d, e Overview, NrCAM can be found in the KIO. lo (d, f) NrCAM can be detected in epithelial cells located in the embryonic cochlear duct at E14. e, g At E17 immunofluorescence images, NrCAM can be observed in the OC. Hes5 staining served as marker for embryonic cochlea. a, c Image by light microscopy, d–g images by immunofluorescence microscopy. Four mice per embryonic day were analyzed. Scale bar=50 μm
Polyclonal Rabbit Antibodies Against Nrcam, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nrcam+concentrations/Mouse%2FRat+NrCAM+Antibody/pm25407819-73-5-11
Average 91 stars, based on 1 article reviews
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Anti-NrCAM (MOUSE) Monoclonal Antibody Allophycocyanin Conjugated - 200-326-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody ATTO 700 Conjugated - 200-362-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody ATTO 565 Conjugated - 200-358-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody ATTO 488 Conjugated - 200-352-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody ATTO 633 Conjugated - 200-360-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody Streptavidin Conjugated - 200-366-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody Biotin Conjugated - 200-306-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody ATTO 594 Conjugated - 200-355-W74
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Anti-NrCAM (MOUSE) Monoclonal Antibody ATTO 655 Conjugated - 200-357-W74
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Image Search Results


Bar graphs and receiver operator curves of Neuronal Cell Adhesion Molecule (NrCAM) in two large cohorts ( a-d , Cohort 1, e-h , Cohort 2). NrCAM concentrations significantly reduced ( a , p = 4.75 x 10 −6 ) in the circulation of women at 36 weeks' gestation who subsequently delivered an FGR infant (birthweight <3rd centile). The discriminatory power of NrCAM is shown as a receiver operating characteristic (AUROC) curve with an area of the curve of 0.76 ( b ). Plasma Placental Growth Factor ( c, d ; PlGF) concentrations at 36 weeks were also significantly reduced in the same cohort ( c , p = 8.15 × 10 −7 ). The AUROC of PlGF is 0.77 ( d ). Cohort, n = 957 controls, n = 26 FGR. In a high-risk international cohort (FEMINA, Manchester, UK) ( e , f ) show NrCAM concentrations reduced ( e , p = 9.34 x 10 −3 ) in the circulation of women who presented at the clinic with reduced fetal movements who subsequently delivered an FGR infant (birthweight <3rd centile), AUROC of 0.72 ( f ). Plasma PlGF ( g, h ) concentrations in the same cohort remain unchanged ( g , p = 0.13). The AUROC of PlGF is 0.63 ( h ). Cohort, n = 235 controls, n = 12 FGR. groups compared using Mann–Whitney U tests. AUROC, area under the AUROC curve, with 95% confidence intervals presented in brackets. Data depicted for a , c, e and g are mean ± SEM. ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

Journal: eBioMedicine

Article Title: Reduced circulating NrCAM as a biomarker for fetal growth restriction

doi: 10.1016/j.ebiom.2025.105854

Figure Lengend Snippet: Bar graphs and receiver operator curves of Neuronal Cell Adhesion Molecule (NrCAM) in two large cohorts ( a-d , Cohort 1, e-h , Cohort 2). NrCAM concentrations significantly reduced ( a , p = 4.75 x 10 −6 ) in the circulation of women at 36 weeks' gestation who subsequently delivered an FGR infant (birthweight <3rd centile). The discriminatory power of NrCAM is shown as a receiver operating characteristic (AUROC) curve with an area of the curve of 0.76 ( b ). Plasma Placental Growth Factor ( c, d ; PlGF) concentrations at 36 weeks were also significantly reduced in the same cohort ( c , p = 8.15 × 10 −7 ). The AUROC of PlGF is 0.77 ( d ). Cohort, n = 957 controls, n = 26 FGR. In a high-risk international cohort (FEMINA, Manchester, UK) ( e , f ) show NrCAM concentrations reduced ( e , p = 9.34 x 10 −3 ) in the circulation of women who presented at the clinic with reduced fetal movements who subsequently delivered an FGR infant (birthweight <3rd centile), AUROC of 0.72 ( f ). Plasma PlGF ( g, h ) concentrations in the same cohort remain unchanged ( g , p = 0.13). The AUROC of PlGF is 0.63 ( h ). Cohort, n = 235 controls, n = 12 FGR. groups compared using Mann–Whitney U tests. AUROC, area under the AUROC curve, with 95% confidence intervals presented in brackets. Data depicted for a , c, e and g are mean ± SEM. ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

Article Snippet: Taqman fast advanced Master Mix (Applied Biosystems) and FAM-labelled Taqman Gene expression Assays (Applies Biosystems) to measure the gene expression of mouse NRCAM (Neuronal Cell Adhesion Molecule, Assay ID: Mm00663607_m1). qRT-PCR was performed on the CFX384 (Bio-Rad) with thermocycling parameters: 95 °C for 20 s, 40 cycles of denaturation for 3 s at 95 °C and 60 °C for 30 s. No product was detected in the non-template control and gene expression of human samples and the hTSCs were normalised to the geometric mean of CYC1 (Cytochrome C1, Assay ID: Hs00357717_m1) and TOP1 (DNA Topoisomerase I, Assay ID: Hs00243257_m1) housekeepers.

Techniques: Clinical Proteomics, MANN-WHITNEY

Circulating Neuronal Cell Adhesion Molecule (NrCAM) ( a , b ) concentrations were not altered ( a , p = 0.09) in the circulation of 24 women at 36 weeks' gestation who subsequently developed preeclampsia (PE) at term, compared with 936 controls. The discriminatory power of NrCAM is shown as a receiver operating characteristic curve with a modest area of the curve (AUROC) of 0.60 ( b ). Plasma Placental Growth Factor ( c, d ; PlGF) concentration is significantly reduced in participants who develop preeclampsia at term ( c , p = 5.45 x 10 −8 ). The AUROC of PlGF is 0.84 ( d ). Controls, n = 959 controls, n = 24 PE. In samples from a case control cohort (PROVE, South Africa), circulating NrCAM was significantly reduced ( e , p = 0.03) in participants with severe preeclampsia. The AUROC of 0.70 ( f ). NrCAM is not altered in participants with eclampsia ( g ), with a modest AUROC of 0.59 ( h ). Control, n = 15 controls, n = 27 preeclampsia (PE), n = 29 eclampsia. Groups compared using Mann–Whitney U tests. AUROC, area under the ROC curve, with 95% confidence intervals presented in brackets. Data depicted for a , c, e and f are mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

Journal: eBioMedicine

Article Title: Reduced circulating NrCAM as a biomarker for fetal growth restriction

doi: 10.1016/j.ebiom.2025.105854

Figure Lengend Snippet: Circulating Neuronal Cell Adhesion Molecule (NrCAM) ( a , b ) concentrations were not altered ( a , p = 0.09) in the circulation of 24 women at 36 weeks' gestation who subsequently developed preeclampsia (PE) at term, compared with 936 controls. The discriminatory power of NrCAM is shown as a receiver operating characteristic curve with a modest area of the curve (AUROC) of 0.60 ( b ). Plasma Placental Growth Factor ( c, d ; PlGF) concentration is significantly reduced in participants who develop preeclampsia at term ( c , p = 5.45 x 10 −8 ). The AUROC of PlGF is 0.84 ( d ). Controls, n = 959 controls, n = 24 PE. In samples from a case control cohort (PROVE, South Africa), circulating NrCAM was significantly reduced ( e , p = 0.03) in participants with severe preeclampsia. The AUROC of 0.70 ( f ). NrCAM is not altered in participants with eclampsia ( g ), with a modest AUROC of 0.59 ( h ). Control, n = 15 controls, n = 27 preeclampsia (PE), n = 29 eclampsia. Groups compared using Mann–Whitney U tests. AUROC, area under the ROC curve, with 95% confidence intervals presented in brackets. Data depicted for a , c, e and f are mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.

Article Snippet: Taqman fast advanced Master Mix (Applied Biosystems) and FAM-labelled Taqman Gene expression Assays (Applies Biosystems) to measure the gene expression of mouse NRCAM (Neuronal Cell Adhesion Molecule, Assay ID: Mm00663607_m1). qRT-PCR was performed on the CFX384 (Bio-Rad) with thermocycling parameters: 95 °C for 20 s, 40 cycles of denaturation for 3 s at 95 °C and 60 °C for 30 s. No product was detected in the non-template control and gene expression of human samples and the hTSCs were normalised to the geometric mean of CYC1 (Cytochrome C1, Assay ID: Hs00357717_m1) and TOP1 (DNA Topoisomerase I, Assay ID: Hs00243257_m1) housekeepers.

Techniques: Clinical Proteomics, Concentration Assay, Control, MANN-WHITNEY

Circulating Neuronal Cell Adhesion Molecule (NrCAM) concentrations were significantly reduced in a case control of participants with preterm fetal growth restriction (FGR, <34 weeks' gestation), compared to gestation-matched controls ( a , p = 0.0003). Gestation-matched controls, n = 20, FGR, n = 23. The discriminatory power of NrCAM is shown as a receiver operating characteristic (AUROC) of 0.82 ( c ). Circulating NrCAM is reduced in participants with preterm preeclampsia (<34 weeks' gestation), compared to gestation-matched controls ( b , p = 0.0003). Gestation-matched controls, n = 20, preeclampsia, n = 41. The AUROC is 0.79 ( d ). NrCAM protein concentrations are reduced in placenta lysates from participants who delivered an FGR infant ( e , p = 0.005). Gestation-matched controls, n = 19, FGR, n = 43. NrCAM protein concentrations are reduced in placenta lysates from participants who are diagnosed with preeclampsia ( f, p = 0.0002), compared to gestation-matched controls. Gestation-matched controls, n = 21, preeclampsia, n = 27. NRCAM mRNA expression is not altered in placenta from participants who delivered an FGR infant ( g ) or diagnosed with preeclampsia ( h ) compared with preterm controls. Gestation-matched controls, n = 17, FGR, n = 63, preeclampsia, n = 78. NFASC mRNA expression is significantly reduced in the placenta from participants who delivered an FGR infant ( i, p = 0.03) or developed preeclampsia at term ( j , p = 0.003), compared with gestation-matched controls. Gestation-matched controls, n = 18, FGR, n = 30, preeclampsia, n = 78. Groups compared using Mann–Whitney U tests. Data presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Journal: eBioMedicine

Article Title: Reduced circulating NrCAM as a biomarker for fetal growth restriction

doi: 10.1016/j.ebiom.2025.105854

Figure Lengend Snippet: Circulating Neuronal Cell Adhesion Molecule (NrCAM) concentrations were significantly reduced in a case control of participants with preterm fetal growth restriction (FGR, <34 weeks' gestation), compared to gestation-matched controls ( a , p = 0.0003). Gestation-matched controls, n = 20, FGR, n = 23. The discriminatory power of NrCAM is shown as a receiver operating characteristic (AUROC) of 0.82 ( c ). Circulating NrCAM is reduced in participants with preterm preeclampsia (<34 weeks' gestation), compared to gestation-matched controls ( b , p = 0.0003). Gestation-matched controls, n = 20, preeclampsia, n = 41. The AUROC is 0.79 ( d ). NrCAM protein concentrations are reduced in placenta lysates from participants who delivered an FGR infant ( e , p = 0.005). Gestation-matched controls, n = 19, FGR, n = 43. NrCAM protein concentrations are reduced in placenta lysates from participants who are diagnosed with preeclampsia ( f, p = 0.0002), compared to gestation-matched controls. Gestation-matched controls, n = 21, preeclampsia, n = 27. NRCAM mRNA expression is not altered in placenta from participants who delivered an FGR infant ( g ) or diagnosed with preeclampsia ( h ) compared with preterm controls. Gestation-matched controls, n = 17, FGR, n = 63, preeclampsia, n = 78. NFASC mRNA expression is significantly reduced in the placenta from participants who delivered an FGR infant ( i, p = 0.03) or developed preeclampsia at term ( j , p = 0.003), compared with gestation-matched controls. Gestation-matched controls, n = 18, FGR, n = 30, preeclampsia, n = 78. Groups compared using Mann–Whitney U tests. Data presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Taqman fast advanced Master Mix (Applied Biosystems) and FAM-labelled Taqman Gene expression Assays (Applies Biosystems) to measure the gene expression of mouse NRCAM (Neuronal Cell Adhesion Molecule, Assay ID: Mm00663607_m1). qRT-PCR was performed on the CFX384 (Bio-Rad) with thermocycling parameters: 95 °C for 20 s, 40 cycles of denaturation for 3 s at 95 °C and 60 °C for 30 s. No product was detected in the non-template control and gene expression of human samples and the hTSCs were normalised to the geometric mean of CYC1 (Cytochrome C1, Assay ID: Hs00357717_m1) and TOP1 (DNA Topoisomerase I, Assay ID: Hs00243257_m1) housekeepers.

Techniques: Control, Expressing, MANN-WHITNEY

NRCAM mRNA expression is reduced following differentiation of cytotrophoblast stem cells to syncytiotrophoblast and extravillous trophoblast (EVT) cells. hTSCs were differentiated into syncytiotrophoblast and EVT cells from 0 h to 96 h. To confirm EVT differentiation, reduction in cytotrophoblast marker TEAD4 ( a , p = 0.006 72 h, p = 0.001 96 h), and increase in EVT marker HLAG ( b , p = 0.02 72 h, p = 0.0002 96 h) mRNA expression was observed. To confirm syncytiotrophoblast differentiation, reduction in cytotrophoblast marker TEAD4 ( d ), and increase in syncytiotrophoblast marker SDC1 ( e ) mRNA expression was observed. NRCAM mRNA expression was reduced in EVT ( c ) and syncytiotrophoblast ( f ) following differentiation. NRCAM mRNA expression was reduced when cytotrophoblast hTSC cells ( g ), but not syncytiotrophoblast cells ( h ) exposed to hypoxia (1% O 2 ), compared to control physiological conditions (normoxia; 8% O 2 ). NRCAM mRNA expression was reduced in primary trophoblast cells ( i ) exposed to hypoxia (1% O 2 ) compared to normoxia (8% O 2 ). mRNA expression was normalized to the geometric mean of housekeeper genes. All experiments were repeated 5 times in triplicate. In primary term human trophoblast cells, NRCAM mRNA expression was significantly reduced in cells exposed to hypoxia (1% O 2 ), compared to normoxic (8% O 2 ) controls ( e , normoxia, n = 6, hypoxia n = 6). For data with two groups, unpaired t-test or a Mann–Whitney (non-parametric) test was used. For 3 or more groups, a one-way ANOVA (parametric) or a Kruskal Wallis (nonparametric) test was used. For tests using multiple comparison tests, data was compared to the control group. Data was presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Journal: eBioMedicine

Article Title: Reduced circulating NrCAM as a biomarker for fetal growth restriction

doi: 10.1016/j.ebiom.2025.105854

Figure Lengend Snippet: NRCAM mRNA expression is reduced following differentiation of cytotrophoblast stem cells to syncytiotrophoblast and extravillous trophoblast (EVT) cells. hTSCs were differentiated into syncytiotrophoblast and EVT cells from 0 h to 96 h. To confirm EVT differentiation, reduction in cytotrophoblast marker TEAD4 ( a , p = 0.006 72 h, p = 0.001 96 h), and increase in EVT marker HLAG ( b , p = 0.02 72 h, p = 0.0002 96 h) mRNA expression was observed. To confirm syncytiotrophoblast differentiation, reduction in cytotrophoblast marker TEAD4 ( d ), and increase in syncytiotrophoblast marker SDC1 ( e ) mRNA expression was observed. NRCAM mRNA expression was reduced in EVT ( c ) and syncytiotrophoblast ( f ) following differentiation. NRCAM mRNA expression was reduced when cytotrophoblast hTSC cells ( g ), but not syncytiotrophoblast cells ( h ) exposed to hypoxia (1% O 2 ), compared to control physiological conditions (normoxia; 8% O 2 ). NRCAM mRNA expression was reduced in primary trophoblast cells ( i ) exposed to hypoxia (1% O 2 ) compared to normoxia (8% O 2 ). mRNA expression was normalized to the geometric mean of housekeeper genes. All experiments were repeated 5 times in triplicate. In primary term human trophoblast cells, NRCAM mRNA expression was significantly reduced in cells exposed to hypoxia (1% O 2 ), compared to normoxic (8% O 2 ) controls ( e , normoxia, n = 6, hypoxia n = 6). For data with two groups, unpaired t-test or a Mann–Whitney (non-parametric) test was used. For 3 or more groups, a one-way ANOVA (parametric) or a Kruskal Wallis (nonparametric) test was used. For tests using multiple comparison tests, data was compared to the control group. Data was presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Taqman fast advanced Master Mix (Applied Biosystems) and FAM-labelled Taqman Gene expression Assays (Applies Biosystems) to measure the gene expression of mouse NRCAM (Neuronal Cell Adhesion Molecule, Assay ID: Mm00663607_m1). qRT-PCR was performed on the CFX384 (Bio-Rad) with thermocycling parameters: 95 °C for 20 s, 40 cycles of denaturation for 3 s at 95 °C and 60 °C for 30 s. No product was detected in the non-template control and gene expression of human samples and the hTSCs were normalised to the geometric mean of CYC1 (Cytochrome C1, Assay ID: Hs00357717_m1) and TOP1 (DNA Topoisomerase I, Assay ID: Hs00243257_m1) housekeepers.

Techniques: Expressing, Marker, Control, MANN-WHITNEY, Comparison

In a mouse model of hypoxia-induced fetal growth restriction, fetal weight ( a ) is significantly reduced in hypoxia treated mothers (10% inspired O 2 ), compared to normoxia controls (21% inspired O 2 ). Placental weight remained unchanged ( b ), whilst placenta-to-body ratio is significantly increased in maternal hypoxia treated group, compared with normoxic controls ( c ). NrCAM mRNA expression is significantly decreased in placentas from the maternal hypoxia group, compared with normoxic controls ( d , n = 9 normoxic placentas, n = 9 hypoxic placentas from separate litters, n = 9 mice in each group). Data was presented as mean ± SEM. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Journal: eBioMedicine

Article Title: Reduced circulating NrCAM as a biomarker for fetal growth restriction

doi: 10.1016/j.ebiom.2025.105854

Figure Lengend Snippet: In a mouse model of hypoxia-induced fetal growth restriction, fetal weight ( a ) is significantly reduced in hypoxia treated mothers (10% inspired O 2 ), compared to normoxia controls (21% inspired O 2 ). Placental weight remained unchanged ( b ), whilst placenta-to-body ratio is significantly increased in maternal hypoxia treated group, compared with normoxic controls ( c ). NrCAM mRNA expression is significantly decreased in placentas from the maternal hypoxia group, compared with normoxic controls ( d , n = 9 normoxic placentas, n = 9 hypoxic placentas from separate litters, n = 9 mice in each group). Data was presented as mean ± SEM. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Taqman fast advanced Master Mix (Applied Biosystems) and FAM-labelled Taqman Gene expression Assays (Applies Biosystems) to measure the gene expression of mouse NRCAM (Neuronal Cell Adhesion Molecule, Assay ID: Mm00663607_m1). qRT-PCR was performed on the CFX384 (Bio-Rad) with thermocycling parameters: 95 °C for 20 s, 40 cycles of denaturation for 3 s at 95 °C and 60 °C for 30 s. No product was detected in the non-template control and gene expression of human samples and the hTSCs were normalised to the geometric mean of CYC1 (Cytochrome C1, Assay ID: Hs00357717_m1) and TOP1 (DNA Topoisomerase I, Assay ID: Hs00243257_m1) housekeepers.

Techniques: Expressing

Fig. 3. Expression and cleavage of selected gelatinase substrates in TNF- α and unstimulated WT astrocyte cultures. (A) Immunofluorescence staining for MMP substrates, VCAM-1, NrCAM, agrin, NOTCH3, together with GFAP to mark astrocytes and DAPI; boxed areas are shown to the right at higher magnifications. Scale bars, 100 μm. (B) Silver-stained gels showing cleavage products of gelatinase substrates after overnight incubation without (0) or with 1:10 or 1:100 ratios of MMP-9:substrate or ADAM10:substrate. Arrows mark the positions of ADAM10 in samples. Asterisks mark specific cleavage products. Data are representative of two to three experiments.

Journal: Science advances

Article Title: Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function.

doi: 10.1126/sciadv.adg0686

Figure Lengend Snippet: Fig. 3. Expression and cleavage of selected gelatinase substrates in TNF- α and unstimulated WT astrocyte cultures. (A) Immunofluorescence staining for MMP substrates, VCAM-1, NrCAM, agrin, NOTCH3, together with GFAP to mark astrocytes and DAPI; boxed areas are shown to the right at higher magnifications. Scale bars, 100 μm. (B) Silver-stained gels showing cleavage products of gelatinase substrates after overnight incubation without (0) or with 1:10 or 1:100 ratios of MMP-9:substrate or ADAM10:substrate. Arrows mark the positions of ADAM10 in samples. Asterisks mark specific cleavage products. Data are representative of two to three experiments.

Article Snippet: To check the ability of recombinant mouse MMP-9 and MMP-2 (R&D Systems) or recombinant mouse ADAM10 (R&D Systems) to in vitro cleave targets identified in the secretome analyses, mouse recombinant VCAM-1 (His-Tag) (Biozol), N-cadherin Fcchimera (R&D Systems), cadherin-4 (R&D Systems), cadherin-11 Fc-chimera (R&D Systems), mouse recombinant NrCAM (R&D Systems), and recombinant rat agrin (R&D Systems) were diluted in 50 mM tris-HCl (pH 7.4), 200 mM NaCl, 5 mM CaCl2, 1 mM APMA, and 0.05% Brij35 to a final concentration of 40 μg/ml, and MMP-9 or MMP-2 was added to a final concentration of 4 μg/ml (10:1 ratio) or 400 ng/ml (100:1 ratio).

Techniques: Expressing, Immunofluorescence, Staining, Incubation

Fig. 4. In vivo expression of MMP substrates, NrCAM and VCAM-1, in noninflamed (naïve) and EAE brains. WT brain sections were immunofluorescently stained for (A) GFAP to mark astrocytes, anti–laminin-γ1 chain antibody to mark BMs and perivascular cuffs, and NrCAM; DAPI marks all nuclei; scale bars, 100 μm; areas marked by the dotted lines are shown at higher magnifications in boxed areas. (B) Immunofluorescence staining for VCAM-1 and pan-laminin (Pan LM) in naïve and in early- and late- stage EAE brains; arrow marks VCAM-1 in CNS parenchyma at late-stage EAE; DAPI marks nuclei; scale bars, 100 μm (naïve) and 50 μm (early/late cuff). (C and D) Triple immunofluorescence staining for CD45, Pan LM, and VCAM-1 shows up-regulation of VCAM-1 around inflammatory cuffs, and its loss at this site where CD45 infiltration occurs (arrows), which correlates with sites of gelatinase activity as shown by in situ zymography (C, bottom) performed on consecutive sections; images to the far right in (C) show in situ hybridizations performed in the presence of the MMP inhibitor, 1,10-phenantrolin. Boxed area in (D) is shown at higher magnification in bottom panels; yellow asterisks mark vessel lumen. Scale bars, 100 μm (C) and 50 μm (D).

Journal: Science advances

Article Title: Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function.

doi: 10.1126/sciadv.adg0686

Figure Lengend Snippet: Fig. 4. In vivo expression of MMP substrates, NrCAM and VCAM-1, in noninflamed (naïve) and EAE brains. WT brain sections were immunofluorescently stained for (A) GFAP to mark astrocytes, anti–laminin-γ1 chain antibody to mark BMs and perivascular cuffs, and NrCAM; DAPI marks all nuclei; scale bars, 100 μm; areas marked by the dotted lines are shown at higher magnifications in boxed areas. (B) Immunofluorescence staining for VCAM-1 and pan-laminin (Pan LM) in naïve and in early- and late- stage EAE brains; arrow marks VCAM-1 in CNS parenchyma at late-stage EAE; DAPI marks nuclei; scale bars, 100 μm (naïve) and 50 μm (early/late cuff). (C and D) Triple immunofluorescence staining for CD45, Pan LM, and VCAM-1 shows up-regulation of VCAM-1 around inflammatory cuffs, and its loss at this site where CD45 infiltration occurs (arrows), which correlates with sites of gelatinase activity as shown by in situ zymography (C, bottom) performed on consecutive sections; images to the far right in (C) show in situ hybridizations performed in the presence of the MMP inhibitor, 1,10-phenantrolin. Boxed area in (D) is shown at higher magnification in bottom panels; yellow asterisks mark vessel lumen. Scale bars, 100 μm (C) and 50 μm (D).

Article Snippet: To check the ability of recombinant mouse MMP-9 and MMP-2 (R&D Systems) or recombinant mouse ADAM10 (R&D Systems) to in vitro cleave targets identified in the secretome analyses, mouse recombinant VCAM-1 (His-Tag) (Biozol), N-cadherin Fcchimera (R&D Systems), cadherin-4 (R&D Systems), cadherin-11 Fc-chimera (R&D Systems), mouse recombinant NrCAM (R&D Systems), and recombinant rat agrin (R&D Systems) were diluted in 50 mM tris-HCl (pH 7.4), 200 mM NaCl, 5 mM CaCl2, 1 mM APMA, and 0.05% Brij35 to a final concentration of 40 μg/ml, and MMP-9 or MMP-2 was added to a final concentration of 4 μg/ml (10:1 ratio) or 400 ng/ml (100:1 ratio).

Techniques: In Vivo, Expressing, Staining, Immunofluorescence, Activity Assay, In Situ, Zymography

Fig. 6. NrCAM function on astrocytes. (A) Immunofluorescence staining of WT and DKO astrocyte-neuronal cocultures for vGlut to mark excitatory synapses and vGAT to mark inhibitory synapses, plus GFAP to mark astrocytes. Wavelet transformations of synapse stainings are shown in bottom panels; scale bars, 20 μm. (B) Corresponding statistical analysis of four experiments with separate culture preparations; data are expressed as relative frequency of GABAergic compared to glutamatergic synapses. Data are means ± SD with two replicates and seven to eight regions analyzed per experiment with each region comprising around 500 to 1000 synapses. Statistical analysis was Student’s t test; *P < 0.05. (C) Immunofluorescence staining of NrCAM, GFAP, and either MAP2 to mark neurons, vGlut, or vGAT in WT and DKO astrocyte- neuronal cocultures; scale bars, 10 μm.

Journal: Science advances

Article Title: Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function.

doi: 10.1126/sciadv.adg0686

Figure Lengend Snippet: Fig. 6. NrCAM function on astrocytes. (A) Immunofluorescence staining of WT and DKO astrocyte-neuronal cocultures for vGlut to mark excitatory synapses and vGAT to mark inhibitory synapses, plus GFAP to mark astrocytes. Wavelet transformations of synapse stainings are shown in bottom panels; scale bars, 20 μm. (B) Corresponding statistical analysis of four experiments with separate culture preparations; data are expressed as relative frequency of GABAergic compared to glutamatergic synapses. Data are means ± SD with two replicates and seven to eight regions analyzed per experiment with each region comprising around 500 to 1000 synapses. Statistical analysis was Student’s t test; *P < 0.05. (C) Immunofluorescence staining of NrCAM, GFAP, and either MAP2 to mark neurons, vGlut, or vGAT in WT and DKO astrocyte- neuronal cocultures; scale bars, 10 μm.

Article Snippet: To check the ability of recombinant mouse MMP-9 and MMP-2 (R&D Systems) or recombinant mouse ADAM10 (R&D Systems) to in vitro cleave targets identified in the secretome analyses, mouse recombinant VCAM-1 (His-Tag) (Biozol), N-cadherin Fcchimera (R&D Systems), cadherin-4 (R&D Systems), cadherin-11 Fc-chimera (R&D Systems), mouse recombinant NrCAM (R&D Systems), and recombinant rat agrin (R&D Systems) were diluted in 50 mM tris-HCl (pH 7.4), 200 mM NaCl, 5 mM CaCl2, 1 mM APMA, and 0.05% Brij35 to a final concentration of 40 μg/ml, and MMP-9 or MMP-2 was added to a final concentration of 4 μg/ml (10:1 ratio) or 400 ng/ml (100:1 ratio).

Techniques: Immunofluorescence, Staining

Fig. 7. Identification of VCAM-1 and NrCAM in multiple sclerosis CSF samples and brain sections. (A) Representative gelatin gel zymography of CSF samples from patients with multiple sclerosis (MS1 to MS6) and age- and sex-matched somatoform controls (details in table S1). NGAL is neutrophil gelatinase–associated lipocalin. (B) ELISA for total MMP-9 in relapsing-remitting multiple sclerosis (RRMS) (n = 15) and somatoform (n = 15) CSF samples; statistical analyses were Mann-Whitney, **P < 0.005. The same CSF samples were tested in (C) ELISA for soluble VCAM-1 (sVCAM-1); data are expressed as change relative to somatoform controls. AU is arbitrary units. (D) Western blot for soluble NrCAM (sNrCAM); bar graph shows quantification of band intensities. Statistical analyses were Student’s t test (C) and Mann-Whitney (D); **P < 0.005 and ****P < 0.0001. (E) Double immunofluorescence staining for GFAP and VCAM-1 or NrCAM in normal-appearing white matter (NAWM), perivascular infiltrates in NAWM, and demyelinating lesions; scale bars, 100 μm. Arrows mark VCAM-1 expressed at the perivascular border and in the CNS parenchyma; bottom panels showing an astrocyte-expressing VCAM-1 in the demyelinating lesion at a higher magnification; scale bar, 25 μm.

Journal: Science advances

Article Title: Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function.

doi: 10.1126/sciadv.adg0686

Figure Lengend Snippet: Fig. 7. Identification of VCAM-1 and NrCAM in multiple sclerosis CSF samples and brain sections. (A) Representative gelatin gel zymography of CSF samples from patients with multiple sclerosis (MS1 to MS6) and age- and sex-matched somatoform controls (details in table S1). NGAL is neutrophil gelatinase–associated lipocalin. (B) ELISA for total MMP-9 in relapsing-remitting multiple sclerosis (RRMS) (n = 15) and somatoform (n = 15) CSF samples; statistical analyses were Mann-Whitney, **P < 0.005. The same CSF samples were tested in (C) ELISA for soluble VCAM-1 (sVCAM-1); data are expressed as change relative to somatoform controls. AU is arbitrary units. (D) Western blot for soluble NrCAM (sNrCAM); bar graph shows quantification of band intensities. Statistical analyses were Student’s t test (C) and Mann-Whitney (D); **P < 0.005 and ****P < 0.0001. (E) Double immunofluorescence staining for GFAP and VCAM-1 or NrCAM in normal-appearing white matter (NAWM), perivascular infiltrates in NAWM, and demyelinating lesions; scale bars, 100 μm. Arrows mark VCAM-1 expressed at the perivascular border and in the CNS parenchyma; bottom panels showing an astrocyte-expressing VCAM-1 in the demyelinating lesion at a higher magnification; scale bar, 25 μm.

Article Snippet: To check the ability of recombinant mouse MMP-9 and MMP-2 (R&D Systems) or recombinant mouse ADAM10 (R&D Systems) to in vitro cleave targets identified in the secretome analyses, mouse recombinant VCAM-1 (His-Tag) (Biozol), N-cadherin Fcchimera (R&D Systems), cadherin-4 (R&D Systems), cadherin-11 Fc-chimera (R&D Systems), mouse recombinant NrCAM (R&D Systems), and recombinant rat agrin (R&D Systems) were diluted in 50 mM tris-HCl (pH 7.4), 200 mM NaCl, 5 mM CaCl2, 1 mM APMA, and 0.05% Brij35 to a final concentration of 40 μg/ml, and MMP-9 or MMP-2 was added to a final concentration of 4 μg/ml (10:1 ratio) or 400 ng/ml (100:1 ratio).

Techniques: Zymography, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Western Blot, Double Immunofluorescence Staining, Expressing

Fig. 1 Immunolabeling of NrCAM in the mouse cochlea at E14 and E17. Cell nuclei are in blue, NrCAM in red, Hes5 in green. a, c Overview of sagittal section of mouse head at the embryonic stage E14 and E17, stained with Hematoxylin/Eosine. Boxed is the cochlea. b The diagram of the embryonic OC. Source: http://syllabus.med.unc. edu. d, e Overview, NrCAM can be found in the KIO. lo (d, f) NrCAM can be detected in epithelial cells located in the embryonic cochlear duct at E14. e, g At E17 immunofluorescence images, NrCAM can be observed in the OC. Hes5 staining served as marker for embryonic cochlea. a, c Image by light microscopy, d–g images by immunofluorescence microscopy. Four mice per embryonic day were analyzed. Scale bar=50 μm

Journal: Journal of molecular neuroscience : MN

Article Title: Neural cell adhesion molecule NrCAM is expressed in the mammalian inner ear and modulates spiral ganglion neurite outgrowth in an in vitro alternate choice assay.

doi: 10.1007/s12031-014-0436-y

Figure Lengend Snippet: Fig. 1 Immunolabeling of NrCAM in the mouse cochlea at E14 and E17. Cell nuclei are in blue, NrCAM in red, Hes5 in green. a, c Overview of sagittal section of mouse head at the embryonic stage E14 and E17, stained with Hematoxylin/Eosine. Boxed is the cochlea. b The diagram of the embryonic OC. Source: http://syllabus.med.unc. edu. d, e Overview, NrCAM can be found in the KIO. lo (d, f) NrCAM can be detected in epithelial cells located in the embryonic cochlear duct at E14. e, g At E17 immunofluorescence images, NrCAM can be observed in the OC. Hes5 staining served as marker for embryonic cochlea. a, c Image by light microscopy, d–g images by immunofluorescence microscopy. Four mice per embryonic day were analyzed. Scale bar=50 μm

Article Snippet: NrCAM stripes were visualized using polyclonal rabbit antibodies against NrCAM (1:400; R&D Systems).

Techniques: Immunolabeling, Staining, Immunofluorescence, Marker, Light Microscopy, Microscopy

Fig. 2 The protein expression pattern of NrCAM in the mouse cochlea from P0 through P21. Cell nuclei are in blue, NrCAM in red, myosin 7A in green served as marker for HCs. a Diagram of the OC. Source: http://syllabus.med. unc.edu. b Negative control. c NrCAM labeling included the region of the IHC, OHC, and DC. d NrCAM labeling was slightly decreased. e Levels of NrCAM labeling is strong in the target zone of the IHC and OHC. f NrCAM labeling is strongest in the region of the OHC. g NrCAM labeling is strong at the IHC and OHC region. b–g Immunofluorescence microscopy images. Four mice per postnatal day were analyzed. Scale bar= 100 μm

Journal: Journal of molecular neuroscience : MN

Article Title: Neural cell adhesion molecule NrCAM is expressed in the mammalian inner ear and modulates spiral ganglion neurite outgrowth in an in vitro alternate choice assay.

doi: 10.1007/s12031-014-0436-y

Figure Lengend Snippet: Fig. 2 The protein expression pattern of NrCAM in the mouse cochlea from P0 through P21. Cell nuclei are in blue, NrCAM in red, myosin 7A in green served as marker for HCs. a Diagram of the OC. Source: http://syllabus.med. unc.edu. b Negative control. c NrCAM labeling included the region of the IHC, OHC, and DC. d NrCAM labeling was slightly decreased. e Levels of NrCAM labeling is strong in the target zone of the IHC and OHC. f NrCAM labeling is strongest in the region of the OHC. g NrCAM labeling is strong at the IHC and OHC region. b–g Immunofluorescence microscopy images. Four mice per postnatal day were analyzed. Scale bar= 100 μm

Article Snippet: NrCAM stripes were visualized using polyclonal rabbit antibodies against NrCAM (1:400; R&D Systems).

Techniques: Expressing, Marker, Negative Control, Labeling, Immunofluorescence, Microscopy

Fig. 3 Quantitative analysis of termination by P5 rat SG neurites on alternating stripes of NrCAM (1, 5, or 10 μg/ml, respectively) versus PLL (5 μg/ml). Neurites showed a dose-dependent preference of NrCAM. Bars represent mean and SD of neurites/explants. *p<0.05, Mann–Whit- ney U test. 16–20 SG explants were analyzed per experimental condition

Journal: Journal of molecular neuroscience : MN

Article Title: Neural cell adhesion molecule NrCAM is expressed in the mammalian inner ear and modulates spiral ganglion neurite outgrowth in an in vitro alternate choice assay.

doi: 10.1007/s12031-014-0436-y

Figure Lengend Snippet: Fig. 3 Quantitative analysis of termination by P5 rat SG neurites on alternating stripes of NrCAM (1, 5, or 10 μg/ml, respectively) versus PLL (5 μg/ml). Neurites showed a dose-dependent preference of NrCAM. Bars represent mean and SD of neurites/explants. *p<0.05, Mann–Whit- ney U test. 16–20 SG explants were analyzed per experimental condition

Article Snippet: NrCAM stripes were visualized using polyclonal rabbit antibodies against NrCAM (1:400; R&D Systems).

Techniques:

Fig. 4 Representative example of a P5 rat SG explant. Explants were cultured on an NrCAM (1, 5, or 10 μg/ml, respectively) versus PLL (5 μg/ml) alternating stripe pattern for 72 h and stained with an anti- neurofilament antibody (red) to visualize all SG neurites, and with an anti-NrCAM antibody (green) to visualize NrCAM. There was a tenden- cy for neurites to terminate upon and/or grow along the NrCAM substrate at NrCAM concentrations of 5 and 10 μg/ml. Stripe width 100 μm

Journal: Journal of molecular neuroscience : MN

Article Title: Neural cell adhesion molecule NrCAM is expressed in the mammalian inner ear and modulates spiral ganglion neurite outgrowth in an in vitro alternate choice assay.

doi: 10.1007/s12031-014-0436-y

Figure Lengend Snippet: Fig. 4 Representative example of a P5 rat SG explant. Explants were cultured on an NrCAM (1, 5, or 10 μg/ml, respectively) versus PLL (5 μg/ml) alternating stripe pattern for 72 h and stained with an anti- neurofilament antibody (red) to visualize all SG neurites, and with an anti-NrCAM antibody (green) to visualize NrCAM. There was a tenden- cy for neurites to terminate upon and/or grow along the NrCAM substrate at NrCAM concentrations of 5 and 10 μg/ml. Stripe width 100 μm

Article Snippet: NrCAM stripes were visualized using polyclonal rabbit antibodies against NrCAM (1:400; R&D Systems).

Techniques: Cell Culture, Staining

Fig. 5 a Effects of NrCAM on the number of neurites extending from SG explants cultivated on increasing concentration of NrCAM. Number of neurites did not differ from control explants grown on PLL 5 μg/ml. Mean of neurites/explant±SD. b Effects of NrCAM on the average length of neurites per SG explant cultivated on increasing concentration of NrCAM. The average length of neurites did not differ from control explants grown on PLL 5 μg/ml. Mean length of neurites/ explant±SD. 16–20 SG explants were analyzed per experimental condition. c Representative example for P5 rat SG explants grown on uniform NrCAM or uniform PLL surface. Explants were cultured for 72 h and stained with an anti-neurofilament primary antibody to visualize all SG neurons

Journal: Journal of molecular neuroscience : MN

Article Title: Neural cell adhesion molecule NrCAM is expressed in the mammalian inner ear and modulates spiral ganglion neurite outgrowth in an in vitro alternate choice assay.

doi: 10.1007/s12031-014-0436-y

Figure Lengend Snippet: Fig. 5 a Effects of NrCAM on the number of neurites extending from SG explants cultivated on increasing concentration of NrCAM. Number of neurites did not differ from control explants grown on PLL 5 μg/ml. Mean of neurites/explant±SD. b Effects of NrCAM on the average length of neurites per SG explant cultivated on increasing concentration of NrCAM. The average length of neurites did not differ from control explants grown on PLL 5 μg/ml. Mean length of neurites/ explant±SD. 16–20 SG explants were analyzed per experimental condition. c Representative example for P5 rat SG explants grown on uniform NrCAM or uniform PLL surface. Explants were cultured for 72 h and stained with an anti-neurofilament primary antibody to visualize all SG neurons

Article Snippet: NrCAM stripes were visualized using polyclonal rabbit antibodies against NrCAM (1:400; R&D Systems).

Techniques: Concentration Assay, Control, Cell Culture, Staining