66687 Search Results


82
Novus Biologicals peroxidase conjugated goat anti human plasminogen
Peroxidase Conjugated Goat Anti Human Plasminogen, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 82/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/peroxidase conjugated goat anti human plasminogen/product/Novus Biologicals
Average 82 stars, based on 1 article reviews
peroxidase conjugated goat anti human plasminogen - by Bioz Stars, 2025-03
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93
Proteintech anti nrbp1 2 antibody
FLAG-HPO-11 interactome a. The difference of log 2 -transformed mean LFQ intensities of FLAG-HPO-11 IP and control IP was plotted against the negative log 10 p-value of a one-sided two-sample t-test (n=3). Proteins with a p-value below 0.05 and a student’s t-test difference of at least 2 were considered as significantly enriched candidates (light grey). The bait protein HPO-11 is shown in green. Homologs of known <t>NRBP1</t> interactors (ELB-1, ELC-1 and Y48C3A.12) are depicted in dark grey, SMG-2 in orange. b. Gene ontology term enrichment analysis for the interactors of HPO-11. Selected enriched GO terms are shown with significance.
Anti Nrbp1 2 Antibody, supplied by Proteintech, 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/result/anti nrbp1 2 antibody/product/Proteintech
Average 93 stars, based on 1 article reviews
anti nrbp1 2 antibody - by Bioz Stars, 2025-03
93/100 stars
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93
Proteintech nrbp2
<t>NRBP2</t> expression is downregulated in BC tissues. (A–C) Relative levels of NRBP2 in BC tissues and adjacent tissues in TCGA database from GEPIA, UCSC Xena and UALCAN. (D) NRBP2 expression in different stages of BC (UALCAN). (E) NRBP2 expression in different subtypes of BC (UALCAN). (F) NRBP2 expression in different lymph node metastasis stages of BC (UALCAN). (G) Levels of the NRBP2 protein in BC tissues and adjacent tissues. (H,I) Immunohistochemical staining for NRBP2 in BC tissues and adjacent tissues (magnification ×200). Quantitative analysis is shown in (I) . * p < 0.05 and *** p < 0.001 compared with the control group.
Nrbp2, supplied by Proteintech, 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/result/nrbp2/product/Proteintech
Average 93 stars, based on 1 article reviews
nrbp2 - by Bioz Stars, 2025-03
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93
Proteintech antibody rabbit anti nrbp2
Expression of nuclear receptor binding protein 2 <t>(NRBP2)</t> in human brain tumors. ( A ) Quantification of the NRBP2 positive staining area in human brain tumors; ( B ) Quantification of NRBP2 staining intensity in human brain tumors; ( C ) Bubble plot comparing the staining quantity (% positive staining) and intensity of NRBP2 in the nucleus (left) or cytoplasm (right) across all samples; ( D ) Immunohistochemistry staining of NRBP2 in non-tumor brain and medulloblastoma tissues of the same tissue microarray, scale bar 20 µm; ( E ) Top panel: NRBP2 protein expression (western blot) in fetal human cerebellum and in medulloblastoma cells D283, D324, MB002 and sD425, Bottom panel: Relative NRBP2 protein expression, normalized to b-actin as loading control of the western blot above, setting fetal human cerebellum (CB) = 1; ( F ) NRBP2 gene expression levels (GSE107405) in MB cell lines (D283, D324, MB002, sD425) in triplicates, and single samples of human cerebellar astrocytes (HA-c) and human spinal cord astrocytes (HA-sp); ( G ) NRBP2 is mainly present in the cytoplasm as seen by subcellular fractionation followed by western blot of cell lysates from a central nervous system(CNS) embryonal tumor cell line PFSK, MB cell lines D283 and D324, and glioblastoma cells U3013MG (Left panel) and the corresponding quantification of intensity (Right panel).
Antibody Rabbit Anti Nrbp2, supplied by Proteintech, 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/result/antibody rabbit anti nrbp2/product/Proteintech
Average 93 stars, based on 1 article reviews
antibody rabbit anti nrbp2 - by Bioz Stars, 2025-03
93/100 stars
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Image Search Results


FLAG-HPO-11 interactome a. The difference of log 2 -transformed mean LFQ intensities of FLAG-HPO-11 IP and control IP was plotted against the negative log 10 p-value of a one-sided two-sample t-test (n=3). Proteins with a p-value below 0.05 and a student’s t-test difference of at least 2 were considered as significantly enriched candidates (light grey). The bait protein HPO-11 is shown in green. Homologs of known NRBP1 interactors (ELB-1, ELC-1 and Y48C3A.12) are depicted in dark grey, SMG-2 in orange. b. Gene ontology term enrichment analysis for the interactors of HPO-11. Selected enriched GO terms are shown with significance.

Journal: bioRxiv

Article Title: Pseudokinase HPO-11 inhibits nonsense-mediated decay to ensure genome stability in C. elegans

doi: 10.1101/2022.09.04.506508

Figure Lengend Snippet: FLAG-HPO-11 interactome a. The difference of log 2 -transformed mean LFQ intensities of FLAG-HPO-11 IP and control IP was plotted against the negative log 10 p-value of a one-sided two-sample t-test (n=3). Proteins with a p-value below 0.05 and a student’s t-test difference of at least 2 were considered as significantly enriched candidates (light grey). The bait protein HPO-11 is shown in green. Homologs of known NRBP1 interactors (ELB-1, ELC-1 and Y48C3A.12) are depicted in dark grey, SMG-2 in orange. b. Gene ontology term enrichment analysis for the interactors of HPO-11. Selected enriched GO terms are shown with significance.

Article Snippet: Anti-Flag (F3165, Sigma-Aldrich), anti-V5-Tag antibody (ab27671, Abcam), anti-GFP antibody (ab290, Abcam), Anti-DNA-RNA hybrid antibody S9.6 (MABE1095, Sigma-Aldrich), anti-NRBP1/2 antibody (21549-1-AP, Proteintech Europe), Phospho-Histone γH2AX antibody (Ser139) (9718, Cell Signaling Technology) and GAPDH (ab8245, Abcam).

Techniques: Transformation Assay

h p o -11 and its human homologs NRBP1 and NRBP2 share conserved functions a. NRBP1/2 interact with UPF1 in MCF-7 cells. Shown is one representative result of three biological replicates. b. siRNA knock-down of either NRBP1 or NRBP2 caused DNA damage response. Shown is a western blot of γ-H2AX in HEK293T cell, N = 3 biological replicates.

Journal: bioRxiv

Article Title: Pseudokinase HPO-11 inhibits nonsense-mediated decay to ensure genome stability in C. elegans

doi: 10.1101/2022.09.04.506508

Figure Lengend Snippet: h p o -11 and its human homologs NRBP1 and NRBP2 share conserved functions a. NRBP1/2 interact with UPF1 in MCF-7 cells. Shown is one representative result of three biological replicates. b. siRNA knock-down of either NRBP1 or NRBP2 caused DNA damage response. Shown is a western blot of γ-H2AX in HEK293T cell, N = 3 biological replicates.

Article Snippet: Anti-Flag (F3165, Sigma-Aldrich), anti-V5-Tag antibody (ab27671, Abcam), anti-GFP antibody (ab290, Abcam), Anti-DNA-RNA hybrid antibody S9.6 (MABE1095, Sigma-Aldrich), anti-NRBP1/2 antibody (21549-1-AP, Proteintech Europe), Phospho-Histone γH2AX antibody (Ser139) (9718, Cell Signaling Technology) and GAPDH (ab8245, Abcam).

Techniques: Western Blot

NRBP2 expression is downregulated in BC tissues. (A–C) Relative levels of NRBP2 in BC tissues and adjacent tissues in TCGA database from GEPIA, UCSC Xena and UALCAN. (D) NRBP2 expression in different stages of BC (UALCAN). (E) NRBP2 expression in different subtypes of BC (UALCAN). (F) NRBP2 expression in different lymph node metastasis stages of BC (UALCAN). (G) Levels of the NRBP2 protein in BC tissues and adjacent tissues. (H,I) Immunohistochemical staining for NRBP2 in BC tissues and adjacent tissues (magnification ×200). Quantitative analysis is shown in (I) . * p < 0.05 and *** p < 0.001 compared with the control group.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: NRBP2 expression is downregulated in BC tissues. (A–C) Relative levels of NRBP2 in BC tissues and adjacent tissues in TCGA database from GEPIA, UCSC Xena and UALCAN. (D) NRBP2 expression in different stages of BC (UALCAN). (E) NRBP2 expression in different subtypes of BC (UALCAN). (F) NRBP2 expression in different lymph node metastasis stages of BC (UALCAN). (G) Levels of the NRBP2 protein in BC tissues and adjacent tissues. (H,I) Immunohistochemical staining for NRBP2 in BC tissues and adjacent tissues (magnification ×200). Quantitative analysis is shown in (I) . * p < 0.05 and *** p < 0.001 compared with the control group.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Expressing, Immunohistochemical staining, Staining

A poor prognosis of patients with BC presenting low NRBP2 expression. The K-M analysis of the (A) OS curve and (B) RFS curve for all patients created using Kaplan-Meier Plotter. (C) The RFS curve for all patients was derived from GEPIA. (D) The RFS curve for patients with basal-like BC, (E) OS curve for patients with Luminal A BC, (F) RFS curve for patients with Luminal A BC, (G) RFS curve for patients with Luminal B BC, (H) RFS curve for patients with HER2-positive BC and (I) RFS curve for patients with lymph node-positive BC were all created using Kaplan-Meier Plotter.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: A poor prognosis of patients with BC presenting low NRBP2 expression. The K-M analysis of the (A) OS curve and (B) RFS curve for all patients created using Kaplan-Meier Plotter. (C) The RFS curve for all patients was derived from GEPIA. (D) The RFS curve for patients with basal-like BC, (E) OS curve for patients with Luminal A BC, (F) RFS curve for patients with Luminal A BC, (G) RFS curve for patients with Luminal B BC, (H) RFS curve for patients with HER2-positive BC and (I) RFS curve for patients with lymph node-positive BC were all created using Kaplan-Meier Plotter.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Expressing, Derivative Assay

Overexpression of NRBP2 inhibits cell proliferation and invasion in vitro . (A) The transfection efficiency of NRBP2 in MCF7 and MDA-MB-231 cells. (B) Cell viability was measured using the CCK-8 assay after NRBP2 overexpression in two BC lines. (C) The invasion of BC cells overexpressing NRBP2 was confirmed using the Transwell assay. The results from the quantitative analysis of the percentage of invading cells are shown (magnification ×200). (D) The knockdown efficiency of NRBP2 in BC cell lines. (E) Cell growth was measured using the CCK-8 assay after NRBP2 knockdown in two BC lines. (F) The invasion of BC cells with knockdown of NRBP2 was determined using Transwell assay. The results from the quantitative analysis the percentage of invading cells are shown (magnification ×200). The values are presented as the means ± SD from three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the control group.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: Overexpression of NRBP2 inhibits cell proliferation and invasion in vitro . (A) The transfection efficiency of NRBP2 in MCF7 and MDA-MB-231 cells. (B) Cell viability was measured using the CCK-8 assay after NRBP2 overexpression in two BC lines. (C) The invasion of BC cells overexpressing NRBP2 was confirmed using the Transwell assay. The results from the quantitative analysis of the percentage of invading cells are shown (magnification ×200). (D) The knockdown efficiency of NRBP2 in BC cell lines. (E) Cell growth was measured using the CCK-8 assay after NRBP2 knockdown in two BC lines. (F) The invasion of BC cells with knockdown of NRBP2 was determined using Transwell assay. The results from the quantitative analysis the percentage of invading cells are shown (magnification ×200). The values are presented as the means ± SD from three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the control group.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Over Expression, In Vitro, Transfection, CCK-8 Assay, Transwell Assay

Overexpression of NRBP2 inhibits the EMT in BC cells in vitro . (A) Levels of the EMT-related proteins E-cadherin, N-cadherin and Snail in NRBP2-knockdown and -overexpressing cells were detected using Western blotting. (B) Quantitative analysis of the optical density ratio of E-cadherin, N-cadherin and Snail levels normalized to β-Actin levels. (C,D) Images of IHC staining for NRBP2, E-cadherin and N-cadherin in BC tissues (magnification ×200). (E) Correlation analysis for of NRBP2 and E-cadherin/N-cadherin. (F) NRBP2 was knocked down in NRBP2-overexpressing cells. Western blotting was performed to detect the levels of EMT-related proteins. The values are presented as the means ± SD from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with the control group.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: Overexpression of NRBP2 inhibits the EMT in BC cells in vitro . (A) Levels of the EMT-related proteins E-cadherin, N-cadherin and Snail in NRBP2-knockdown and -overexpressing cells were detected using Western blotting. (B) Quantitative analysis of the optical density ratio of E-cadherin, N-cadherin and Snail levels normalized to β-Actin levels. (C,D) Images of IHC staining for NRBP2, E-cadherin and N-cadherin in BC tissues (magnification ×200). (E) Correlation analysis for of NRBP2 and E-cadherin/N-cadherin. (F) NRBP2 was knocked down in NRBP2-overexpressing cells. Western blotting was performed to detect the levels of EMT-related proteins. The values are presented as the means ± SD from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with the control group.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Over Expression, In Vitro, Western Blot, Immunohistochemistry

Overexpression of NRBP2 decreased the lung metastasis of BC cells in vivo . (A) The transfection efficiency of the NRBP2 overexpression plasmid in MDA-MB-231 cells. (B) The efficiency of NRBP2 overexpression in MDA-MB-231 cells. (C) Schematic of the experimental procedure used to assess lung metastasis. (D) Comparison of tumor volume from various groups. (E) Bright field images of the lung metastases in the control group and overexpression group (left panel) and the quantification of the metastatic tumors (right) ( n =4). (F) HE staining of lung metastatic tumors. (magnification ×200) ** p < 0.01 compared with the control group.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: Overexpression of NRBP2 decreased the lung metastasis of BC cells in vivo . (A) The transfection efficiency of the NRBP2 overexpression plasmid in MDA-MB-231 cells. (B) The efficiency of NRBP2 overexpression in MDA-MB-231 cells. (C) Schematic of the experimental procedure used to assess lung metastasis. (D) Comparison of tumor volume from various groups. (E) Bright field images of the lung metastases in the control group and overexpression group (left panel) and the quantification of the metastatic tumors (right) ( n =4). (F) HE staining of lung metastatic tumors. (magnification ×200) ** p < 0.01 compared with the control group.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Over Expression, In Vivo, Transfection, Plasmid Preparation, Staining

NRBP2 regulated the EMT of BC cells via AMPK signaling. (A) Levels of the p-AMPK, AMPK, p-mTOR and mTOR proteins in NRBP2-knockdown and -overexpressing cells were detected using Western blotting. (B) Quantitative analysis of the optical density ratio of p-AMPK, AMPK, p-mTOR and mTOR to β-Actin. (C) The levels of EMT-related proteins in NRBP2-overexpressing cells treated with Com.C (10 μM, 24 h) were detected using Western blotting. (D) Cell viability was measured using the CCK-8 assay in BC cells treated as described above. (E,F) Transwell assays were used to detect the invasion of two BC cell lines treated as described above. (magnification ×200) Right: Quantitative analysis of the percentage of invading cells. The values are presented as the means ± SD from three independent experiments. # p < 0.05, ## p < 0.01, * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with the corresponding group.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: NRBP2 regulated the EMT of BC cells via AMPK signaling. (A) Levels of the p-AMPK, AMPK, p-mTOR and mTOR proteins in NRBP2-knockdown and -overexpressing cells were detected using Western blotting. (B) Quantitative analysis of the optical density ratio of p-AMPK, AMPK, p-mTOR and mTOR to β-Actin. (C) The levels of EMT-related proteins in NRBP2-overexpressing cells treated with Com.C (10 μM, 24 h) were detected using Western blotting. (D) Cell viability was measured using the CCK-8 assay in BC cells treated as described above. (E,F) Transwell assays were used to detect the invasion of two BC cell lines treated as described above. (magnification ×200) Right: Quantitative analysis of the percentage of invading cells. The values are presented as the means ± SD from three independent experiments. # p < 0.05, ## p < 0.01, * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with the corresponding group.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Western Blot, CCK-8 Assay

NRBP2 regulated the EMT in BC cells via AMPK/mTOR signaling. (A) The levels of EMT-related proteins in NRBP2-silenced cells treated with Com.C (1 μM, 24 h) were detected using Western blotting. (B) Cell viability was measured using the CCK-8 assay as described above. (C,D) The Transwell assay revealed the invasion of BC cells treated as described above. (magnification ×200) Right: Quantitative analysis of the percentage of invading cells. (E) Schematic model of the mechanism by which NRBP2 inhibits the progression of BC. The values are presented as the means ± SD from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with the corresponding group.

Journal: Frontiers in Oncology

Article Title: NRBP2 Functions as a Tumor Suppressor and Inhibits Epithelial-to-Mesenchymal Transition in Breast Cancer

doi: 10.3389/fonc.2021.634026

Figure Lengend Snippet: NRBP2 regulated the EMT in BC cells via AMPK/mTOR signaling. (A) The levels of EMT-related proteins in NRBP2-silenced cells treated with Com.C (1 μM, 24 h) were detected using Western blotting. (B) Cell viability was measured using the CCK-8 assay as described above. (C,D) The Transwell assay revealed the invasion of BC cells treated as described above. (magnification ×200) Right: Quantitative analysis of the percentage of invading cells. (E) Schematic model of the mechanism by which NRBP2 inhibits the progression of BC. The values are presented as the means ± SD from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 compared with the corresponding group.

Article Snippet: Antibodies against E-cadherin (Santa Cruz, sc-7870), N-cadherin (Cell Signaling Technology, 13116), Snail (Santa Cruz, sc-393172), NRBP2 (Proteintech, 21549-1-AP), p-AMPK (Cell Signaling Technology, 2535), AMPK (Cell Signaling Technology, 5831), p-mTOR (Cell Signaling Technology, 5536), mTOR (Cell Signaling Technology, 2983), and actin (Sigma, A5441) were used.

Techniques: Western Blot, CCK-8 Assay, Transwell Assay

Expression of nuclear receptor binding protein 2 (NRBP2) in human brain tumors. ( A ) Quantification of the NRBP2 positive staining area in human brain tumors; ( B ) Quantification of NRBP2 staining intensity in human brain tumors; ( C ) Bubble plot comparing the staining quantity (% positive staining) and intensity of NRBP2 in the nucleus (left) or cytoplasm (right) across all samples; ( D ) Immunohistochemistry staining of NRBP2 in non-tumor brain and medulloblastoma tissues of the same tissue microarray, scale bar 20 µm; ( E ) Top panel: NRBP2 protein expression (western blot) in fetal human cerebellum and in medulloblastoma cells D283, D324, MB002 and sD425, Bottom panel: Relative NRBP2 protein expression, normalized to b-actin as loading control of the western blot above, setting fetal human cerebellum (CB) = 1; ( F ) NRBP2 gene expression levels (GSE107405) in MB cell lines (D283, D324, MB002, sD425) in triplicates, and single samples of human cerebellar astrocytes (HA-c) and human spinal cord astrocytes (HA-sp); ( G ) NRBP2 is mainly present in the cytoplasm as seen by subcellular fractionation followed by western blot of cell lysates from a central nervous system(CNS) embryonal tumor cell line PFSK, MB cell lines D283 and D324, and glioblastoma cells U3013MG (Left panel) and the corresponding quantification of intensity (Right panel).

Journal: Cancers

Article Title: Nuclear Receptor Binding Protein 2 Is Downregulated in Medulloblastoma, and Reduces Tumor Cell Survival upon Overexpression

doi: 10.3390/cancers12061483

Figure Lengend Snippet: Expression of nuclear receptor binding protein 2 (NRBP2) in human brain tumors. ( A ) Quantification of the NRBP2 positive staining area in human brain tumors; ( B ) Quantification of NRBP2 staining intensity in human brain tumors; ( C ) Bubble plot comparing the staining quantity (% positive staining) and intensity of NRBP2 in the nucleus (left) or cytoplasm (right) across all samples; ( D ) Immunohistochemistry staining of NRBP2 in non-tumor brain and medulloblastoma tissues of the same tissue microarray, scale bar 20 µm; ( E ) Top panel: NRBP2 protein expression (western blot) in fetal human cerebellum and in medulloblastoma cells D283, D324, MB002 and sD425, Bottom panel: Relative NRBP2 protein expression, normalized to b-actin as loading control of the western blot above, setting fetal human cerebellum (CB) = 1; ( F ) NRBP2 gene expression levels (GSE107405) in MB cell lines (D283, D324, MB002, sD425) in triplicates, and single samples of human cerebellar astrocytes (HA-c) and human spinal cord astrocytes (HA-sp); ( G ) NRBP2 is mainly present in the cytoplasm as seen by subcellular fractionation followed by western blot of cell lysates from a central nervous system(CNS) embryonal tumor cell line PFSK, MB cell lines D283 and D324, and glioblastoma cells U3013MG (Left panel) and the corresponding quantification of intensity (Right panel).

Article Snippet: The membrane was blocked in 5% non-fat milk in TBST (Tris buffered saline supplemented with Tween-20)for 1 h at room temperature and then incubated with primary antibody rabbit anti-NRBP2 (1:1000, Proteintech, Manchester, UK), p-AKT pSer473 (1:000, Cell Signaling Technology, Danvers, MA, USA), cleaved caspase 3 (1:1000, Cell Signaling, MA, USA), mouse monoclonal BAK1 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal Bax (1:500, Santa Cruz Biotechnology), β-Actin (1:500, Santa Cruz Biotechnology), mouse monoclonal Ki67 (1:400, Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: Expressing, Binding Assay, Staining, Immunohistochemistry, Microarray, Western Blot, Fractionation

Expression of nuclear receptor binding protein (NRBP2) in human medulloblastoma. ( A ) NRBP2 gene expression levels (GSE124814) in normal human cerebellum, either for pediatric cases only (age < 18 years; n = 47) or across pediatric and adult cases (all ages; n = 291), and MB patients ( n = 1350); ( B ) NRBP2 gene expression levels (GSE85217) in wingless (WNT) ( n = 70), sonic hedgehog (SHH) ( n = 223), Group 3 ( n = 144), and Group 4 ( n = 326) subgroups of medulloblastoma patients. Boxes indicate the range between the first and third quartiles, black horizontal lines represent the median value, and whiskers extend to the extreme values excluding outliers, which are shown as dots; ( C ) NRBP2 protein expression levels (MSV00008264) in WNT ( n = 3), SHH ( n = 15), Group 3 ( n = 14) and Group 4 ( n = 13) subgroups of MB patients. Boxes indicate the range between 1–99 percentile, black horizontal lines represent the median value, and whiskers extend to the extreme values. Unpaired t test between the SHH and G3 groups showed a significant difference (* denotes significance as p < 0.05).

Journal: Cancers

Article Title: Nuclear Receptor Binding Protein 2 Is Downregulated in Medulloblastoma, and Reduces Tumor Cell Survival upon Overexpression

doi: 10.3390/cancers12061483

Figure Lengend Snippet: Expression of nuclear receptor binding protein (NRBP2) in human medulloblastoma. ( A ) NRBP2 gene expression levels (GSE124814) in normal human cerebellum, either for pediatric cases only (age < 18 years; n = 47) or across pediatric and adult cases (all ages; n = 291), and MB patients ( n = 1350); ( B ) NRBP2 gene expression levels (GSE85217) in wingless (WNT) ( n = 70), sonic hedgehog (SHH) ( n = 223), Group 3 ( n = 144), and Group 4 ( n = 326) subgroups of medulloblastoma patients. Boxes indicate the range between the first and third quartiles, black horizontal lines represent the median value, and whiskers extend to the extreme values excluding outliers, which are shown as dots; ( C ) NRBP2 protein expression levels (MSV00008264) in WNT ( n = 3), SHH ( n = 15), Group 3 ( n = 14) and Group 4 ( n = 13) subgroups of MB patients. Boxes indicate the range between 1–99 percentile, black horizontal lines represent the median value, and whiskers extend to the extreme values. Unpaired t test between the SHH and G3 groups showed a significant difference (* denotes significance as p < 0.05).

Article Snippet: The membrane was blocked in 5% non-fat milk in TBST (Tris buffered saline supplemented with Tween-20)for 1 h at room temperature and then incubated with primary antibody rabbit anti-NRBP2 (1:1000, Proteintech, Manchester, UK), p-AKT pSer473 (1:000, Cell Signaling Technology, Danvers, MA, USA), cleaved caspase 3 (1:1000, Cell Signaling, MA, USA), mouse monoclonal BAK1 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal Bax (1:500, Santa Cruz Biotechnology), β-Actin (1:500, Santa Cruz Biotechnology), mouse monoclonal Ki67 (1:400, Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: Expressing, Binding Assay

Effects on nuclear receptor binding protein (NRBP2) mRNA expression and cell growth of medulloblastoma (MB) cell lines by promotor de-methylation and histone de-acetylation treatment. ( A ) relative NRBP2 expression in D283, D324, sD425, and MB002 cell lines treated with 5-Aza-2′-deoxycytidine (dAC) for two days, expressed as RQ value, 2-(ΔΔCt) value, beta-actin was used as an endogenous control and untreated cells as the reference. n = 3; ( B ) relative NRBP2 expression in D283, D324, sD425, and MB002 cell lines treated with Valproic acid (VPA) for two days, expressed as RQ (relative quantification) value, 2-(ΔΔCt) value, beta-actin was used as an endogenous control and untreated cells as the reference. n = 3; ( C ) Left Upper panel: NRBP2 protein expression in MB002, sD425, and D324 cell lines treated with dAC for two days. Left Lower panel: Signal intensity quantification by ImageJ. Right Upper panel: NRBP2 protein expression in MB002, sD425 and D324 cell lines treated with VPA for two days. Right Lower panel: Signal intensity quantification by ImageJ (an open source image processing program developed at the National Institute of Health, USA) . b-actin was used as a loading control for both; ( D ) Left panel: growth curves for MB002 (upper) and sD425 (lower) cell lines treated with VPA compared to untreated cells. Right panel: growth curves for MB002 (upper) and sD425 (lower) cell lines treated with dAC compared to untreated cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ( E ) Left panel: relative gene expression levels of NCOR after transfection by off-target-siRNA, NCOR-siRNA, SMRT-siRNA, and NCOR and SMRT-siRNA; middle panel: relative gene expression levels of SMRT after treatment by off-target-siRNA, NCOR-siRNA, SMRT-siRNA, and NCOR and SMRT-siRNA; right panel: relative gene expression levels of NRBP2 after treatment by off-target-siRNA, NCOR-siRNA, SMRT-siRNA, and NCOR and SMRT-siRNA. n = 3. Average expression levels after off-target-siRNA transfection were used as control.

Journal: Cancers

Article Title: Nuclear Receptor Binding Protein 2 Is Downregulated in Medulloblastoma, and Reduces Tumor Cell Survival upon Overexpression

doi: 10.3390/cancers12061483

Figure Lengend Snippet: Effects on nuclear receptor binding protein (NRBP2) mRNA expression and cell growth of medulloblastoma (MB) cell lines by promotor de-methylation and histone de-acetylation treatment. ( A ) relative NRBP2 expression in D283, D324, sD425, and MB002 cell lines treated with 5-Aza-2′-deoxycytidine (dAC) for two days, expressed as RQ value, 2-(ΔΔCt) value, beta-actin was used as an endogenous control and untreated cells as the reference. n = 3; ( B ) relative NRBP2 expression in D283, D324, sD425, and MB002 cell lines treated with Valproic acid (VPA) for two days, expressed as RQ (relative quantification) value, 2-(ΔΔCt) value, beta-actin was used as an endogenous control and untreated cells as the reference. n = 3; ( C ) Left Upper panel: NRBP2 protein expression in MB002, sD425, and D324 cell lines treated with dAC for two days. Left Lower panel: Signal intensity quantification by ImageJ. Right Upper panel: NRBP2 protein expression in MB002, sD425 and D324 cell lines treated with VPA for two days. Right Lower panel: Signal intensity quantification by ImageJ (an open source image processing program developed at the National Institute of Health, USA) . b-actin was used as a loading control for both; ( D ) Left panel: growth curves for MB002 (upper) and sD425 (lower) cell lines treated with VPA compared to untreated cells. Right panel: growth curves for MB002 (upper) and sD425 (lower) cell lines treated with dAC compared to untreated cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ( E ) Left panel: relative gene expression levels of NCOR after transfection by off-target-siRNA, NCOR-siRNA, SMRT-siRNA, and NCOR and SMRT-siRNA; middle panel: relative gene expression levels of SMRT after treatment by off-target-siRNA, NCOR-siRNA, SMRT-siRNA, and NCOR and SMRT-siRNA; right panel: relative gene expression levels of NRBP2 after treatment by off-target-siRNA, NCOR-siRNA, SMRT-siRNA, and NCOR and SMRT-siRNA. n = 3. Average expression levels after off-target-siRNA transfection were used as control.

Article Snippet: The membrane was blocked in 5% non-fat milk in TBST (Tris buffered saline supplemented with Tween-20)for 1 h at room temperature and then incubated with primary antibody rabbit anti-NRBP2 (1:1000, Proteintech, Manchester, UK), p-AKT pSer473 (1:000, Cell Signaling Technology, Danvers, MA, USA), cleaved caspase 3 (1:1000, Cell Signaling, MA, USA), mouse monoclonal BAK1 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal Bax (1:500, Santa Cruz Biotechnology), β-Actin (1:500, Santa Cruz Biotechnology), mouse monoclonal Ki67 (1:400, Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: Binding Assay, Expressing, Methylation, Transfection

Overexpression of nuclear receptor binding protein (NRBP2) in D324 medulloblastoma (MB) cells causes reduction of cell numbers, migration, and invasion. ( A ) Western blot of NRBP2 in D324 cells transfected with an empty vector (cntl) or NRBP2-IRES2-eGFP NRBP2 sequence with V5 tag and linked with the internal ribosome entry site (IRES2) and the enhanced green fluorescent protein (eGFP) coding region) overexpression plasmid, Left panel: Signal intensity quantification by ImageJ; ( B ) growth rate of D324 cells transfected with control vector or NRBP2 vector; ( C ) Western blot of NRBP2 in D324 transduced with scrambled shRNA, or shRNA against NRBP2. The cultures were transduced twice to increase the efficiency of downregulation, Left panel: Signal intensity quantification by ImageJ; ( D ) growth curve of D324 cells transduced with scrambled shRNA, or shRNA against NRBP2; ( E ) photomicrographs of cell migration after scratching the cell monolayer ( n = 3); ( F ) quantification of coverage of cell-free area from ( C ) ( n = 3); ( G ) photomicrographs of cell invasion in collagen gel at different time points of D324 cells transduced with scrambled shRNA, or shRNA against NRBP2; ( H ) quantification of invasion from ( E ) ( n = 3). * p < 0.05, ** p < 0.01, **** p < 0.0001.

Journal: Cancers

Article Title: Nuclear Receptor Binding Protein 2 Is Downregulated in Medulloblastoma, and Reduces Tumor Cell Survival upon Overexpression

doi: 10.3390/cancers12061483

Figure Lengend Snippet: Overexpression of nuclear receptor binding protein (NRBP2) in D324 medulloblastoma (MB) cells causes reduction of cell numbers, migration, and invasion. ( A ) Western blot of NRBP2 in D324 cells transfected with an empty vector (cntl) or NRBP2-IRES2-eGFP NRBP2 sequence with V5 tag and linked with the internal ribosome entry site (IRES2) and the enhanced green fluorescent protein (eGFP) coding region) overexpression plasmid, Left panel: Signal intensity quantification by ImageJ; ( B ) growth rate of D324 cells transfected with control vector or NRBP2 vector; ( C ) Western blot of NRBP2 in D324 transduced with scrambled shRNA, or shRNA against NRBP2. The cultures were transduced twice to increase the efficiency of downregulation, Left panel: Signal intensity quantification by ImageJ; ( D ) growth curve of D324 cells transduced with scrambled shRNA, or shRNA against NRBP2; ( E ) photomicrographs of cell migration after scratching the cell monolayer ( n = 3); ( F ) quantification of coverage of cell-free area from ( C ) ( n = 3); ( G ) photomicrographs of cell invasion in collagen gel at different time points of D324 cells transduced with scrambled shRNA, or shRNA against NRBP2; ( H ) quantification of invasion from ( E ) ( n = 3). * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: The membrane was blocked in 5% non-fat milk in TBST (Tris buffered saline supplemented with Tween-20)for 1 h at room temperature and then incubated with primary antibody rabbit anti-NRBP2 (1:1000, Proteintech, Manchester, UK), p-AKT pSer473 (1:000, Cell Signaling Technology, Danvers, MA, USA), cleaved caspase 3 (1:1000, Cell Signaling, MA, USA), mouse monoclonal BAK1 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal Bax (1:500, Santa Cruz Biotechnology), β-Actin (1:500, Santa Cruz Biotechnology), mouse monoclonal Ki67 (1:400, Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: Over Expression, Binding Assay, Migration, Western Blot, Transfection, Plasmid Preparation, Sequencing, Transduction, shRNA

Overexpression of nuclear receptor binding protein (NRBP2) in medulloblastoma (MB) leads to increased cell death. ( A ) flow-cytometry-based quantification of live and dead D324 cells 1 and 2 days post-transfection with NRBP2 -V5-IRES-eGFP (NRBP2 sequence with V5 tag, linked with the internal ribosome entry site (IRES2) and the enhanced green fluorescent protein (eGFP) coding region) plasmid; ( B ) flow-cytometry-based cell-cycle analysis in D324 cells after transfection with NRBP2 -V5-IRES-eGFP plasmid; ( C ) Western blot of Ki67 in MB002 and sD425 cells 2–24 h post transduction with empty vector or EF1A-NRBP2(V5)-plasmid. Corresponding Signal intensity quantification by ImageJ on the left; ( D ) Western blot of phosphorylation of AKT in D324 cells transfected with control or NRBP2 plasmid and signal intensity quantification by ImageJ on the left; ( E ) apoptosis analysis based on Annexin V labelling of control-transfected or NRBP2-transfected D324 cells; ( F ) Western blot of cleaved caspase-3 in MB002 and sD425 cells transduced with empty vector or NRBP2 overexpressing plasmid, 48 h post transduction; ( G ) cytospin preparations of MB002 cells, 4 to 48 h after transduction with a EF1A-NRBP2(V5)- plasmid showing staining for NRBP2 (upper panel) and cleaved caspase-3 (lower panel); ( H ) expression of apoptotic genes BAK1 and BAX mRNA in D324 cells transfected with control or NRBP2 plasmid; ( I ) Western blot of BAK1 and BAX in MB002 and sD425 cells transduced with control or NRBP2 plasmid 48 h post transduction. Corresponding signal intensity quantification by ImageJ is shown on the left.

Journal: Cancers

Article Title: Nuclear Receptor Binding Protein 2 Is Downregulated in Medulloblastoma, and Reduces Tumor Cell Survival upon Overexpression

doi: 10.3390/cancers12061483

Figure Lengend Snippet: Overexpression of nuclear receptor binding protein (NRBP2) in medulloblastoma (MB) leads to increased cell death. ( A ) flow-cytometry-based quantification of live and dead D324 cells 1 and 2 days post-transfection with NRBP2 -V5-IRES-eGFP (NRBP2 sequence with V5 tag, linked with the internal ribosome entry site (IRES2) and the enhanced green fluorescent protein (eGFP) coding region) plasmid; ( B ) flow-cytometry-based cell-cycle analysis in D324 cells after transfection with NRBP2 -V5-IRES-eGFP plasmid; ( C ) Western blot of Ki67 in MB002 and sD425 cells 2–24 h post transduction with empty vector or EF1A-NRBP2(V5)-plasmid. Corresponding Signal intensity quantification by ImageJ on the left; ( D ) Western blot of phosphorylation of AKT in D324 cells transfected with control or NRBP2 plasmid and signal intensity quantification by ImageJ on the left; ( E ) apoptosis analysis based on Annexin V labelling of control-transfected or NRBP2-transfected D324 cells; ( F ) Western blot of cleaved caspase-3 in MB002 and sD425 cells transduced with empty vector or NRBP2 overexpressing plasmid, 48 h post transduction; ( G ) cytospin preparations of MB002 cells, 4 to 48 h after transduction with a EF1A-NRBP2(V5)- plasmid showing staining for NRBP2 (upper panel) and cleaved caspase-3 (lower panel); ( H ) expression of apoptotic genes BAK1 and BAX mRNA in D324 cells transfected with control or NRBP2 plasmid; ( I ) Western blot of BAK1 and BAX in MB002 and sD425 cells transduced with control or NRBP2 plasmid 48 h post transduction. Corresponding signal intensity quantification by ImageJ is shown on the left.

Article Snippet: The membrane was blocked in 5% non-fat milk in TBST (Tris buffered saline supplemented with Tween-20)for 1 h at room temperature and then incubated with primary antibody rabbit anti-NRBP2 (1:1000, Proteintech, Manchester, UK), p-AKT pSer473 (1:000, Cell Signaling Technology, Danvers, MA, USA), cleaved caspase 3 (1:1000, Cell Signaling, MA, USA), mouse monoclonal BAK1 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal Bax (1:500, Santa Cruz Biotechnology), β-Actin (1:500, Santa Cruz Biotechnology), mouse monoclonal Ki67 (1:400, Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: Over Expression, Binding Assay, Flow Cytometry, Transfection, Sequencing, Plasmid Preparation, Cell Cycle Assay, Western Blot, Transduction, Staining, Expressing

List of sequences of the primers used for qPCR.

Journal: Cancers

Article Title: Nuclear Receptor Binding Protein 2 Is Downregulated in Medulloblastoma, and Reduces Tumor Cell Survival upon Overexpression

doi: 10.3390/cancers12061483

Figure Lengend Snippet: List of sequences of the primers used for qPCR.

Article Snippet: The membrane was blocked in 5% non-fat milk in TBST (Tris buffered saline supplemented with Tween-20)for 1 h at room temperature and then incubated with primary antibody rabbit anti-NRBP2 (1:1000, Proteintech, Manchester, UK), p-AKT pSer473 (1:000, Cell Signaling Technology, Danvers, MA, USA), cleaved caspase 3 (1:1000, Cell Signaling, MA, USA), mouse monoclonal BAK1 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal Bax (1:500, Santa Cruz Biotechnology), β-Actin (1:500, Santa Cruz Biotechnology), mouse monoclonal Ki67 (1:400, Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: