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Santa Cruz Biotechnology nrp2 antibody

<t>NRP2</t> expression is up-regulated in osteosarcoma cells. (A) , mRNA levels of NRP2 in normal osteoblast (NHOst) and osteosarcoma cell lines (Saos-LM7, 143B, 143.98.2, MG-63, MNNG/HOS, Saos-2, and U2-OS) were determined by real time PCR. (B) , the protein levels of NRP2 in NHOst and seven osteosarcoma cell lines were detected by western blot using NRP2 antibody. The relative protein levels were determined by densitometry and normalized with β-actin level. (C) , Kaplan-Meier survival curves of disease–specific mortality for patients whose osteosarcoma expressed or didn’t express NRP2. The log-rank test was used to compare differences between two groups. NRP2 expression was predictive of poor overall survival. Significant difference is indicated by (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Column: mean value; Error bars: SEM

Nrp2 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/immunohistochemistry+microarray+based+expression+data/neuropilin-2+Antibody/pmc04411772-221-33-35
Average 93 stars, based on 105 article reviews

nrp2 antibody - by Bioz Stars, 2026-08

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1) Product Images from "Neuropilin-2 expression is inhibited by secreted Wnt antagonists and its down-regulation is associated with reduced tumor growth and metastasis in osteosarcoma"

Article Title: Neuropilin-2 expression is inhibited by secreted Wnt antagonists and its down-regulation is associated with reduced tumor growth and metastasis in osteosarcoma

Journal: Molecular Cancer

doi: 10.1186/s12943-015-0359-4

NRP2 expression is up-regulated in osteosarcoma cells. (A) , mRNA levels of NRP2 in normal osteoblast (NHOst) and osteosarcoma cell lines (Saos-LM7, 143B, 143.98.2, MG-63, MNNG/HOS, Saos-2, and U2-OS) were determined by real time PCR. (B) , the protein levels of NRP2 in NHOst and seven osteosarcoma cell lines were detected by western blot using NRP2 antibody. The relative protein levels were determined by densitometry and normalized with β-actin level. (C) , Kaplan-Meier survival curves of disease–specific mortality for patients whose osteosarcoma expressed or didn’t express NRP2. The log-rank test was used to compare differences between two groups. NRP2 expression was predictive of poor overall survival. Significant difference is indicated by (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Column: mean value; Error bars: SEM

Figure Legend Snippet: NRP2 expression is up-regulated in osteosarcoma cells. (A) , mRNA levels of NRP2 in normal osteoblast (NHOst) and osteosarcoma cell lines (Saos-LM7, 143B, 143.98.2, MG-63, MNNG/HOS, Saos-2, and U2-OS) were determined by real time PCR. (B) , the protein levels of NRP2 in NHOst and seven osteosarcoma cell lines were detected by western blot using NRP2 antibody. The relative protein levels were determined by densitometry and normalized with β-actin level. (C) , Kaplan-Meier survival curves of disease–specific mortality for patients whose osteosarcoma expressed or didn’t express NRP2. The log-rank test was used to compare differences between two groups. NRP2 expression was predictive of poor overall survival. Significant difference is indicated by (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Column: mean value; Error bars: SEM

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot

NRP2 knockdown inhibited both in vitro and in vivo tumor growth. (A & B) , NRP2 expression was knocked down by NRP2 shRNA in 143B cells. Knockdown efficiency was determined by real-time PCR (A) and Western blotting (B) . (C) , By MTT assay, NRP2 knockdown suppressed anchorage-dependent growth of osteosarcoma 143B cells. (D) Soft agar assay. NRP2 knockdown did not reduce the number of colony formed by 143B cells. 143B cells transfected with ShNRP2 formed smaller colony than control vector transfected cells as shown in the representative images of soft agar (insert). (E) & (F) NRP2 knockdown inhibited the in vivo tumor growth in xenograft nude mice model. 1 × 10 6 143B cells were inoculated in NCR nu-nu nude mice. Tumor size was measured every 3 days and a tumor growth curve was created (E) . Points, mean tumor volume; Bars, SEM. (F) , Representative picture of tumor harvested at day 21. (G) , the knockdown of NRP2 expression in tumor samples was confirmed by immunofluorescence staining using NRP2 antibody. Significant differences are indicated by: (*) p < 0.05, (**) p < 0.01. Column: mean value; Error bars: SEM.

Figure Legend Snippet: NRP2 knockdown inhibited both in vitro and in vivo tumor growth. (A & B) , NRP2 expression was knocked down by NRP2 shRNA in 143B cells. Knockdown efficiency was determined by real-time PCR (A) and Western blotting (B) . (C) , By MTT assay, NRP2 knockdown suppressed anchorage-dependent growth of osteosarcoma 143B cells. (D) Soft agar assay. NRP2 knockdown did not reduce the number of colony formed by 143B cells. 143B cells transfected with ShNRP2 formed smaller colony than control vector transfected cells as shown in the representative images of soft agar (insert). (E) & (F) NRP2 knockdown inhibited the in vivo tumor growth in xenograft nude mice model. 1 × 10 6 143B cells were inoculated in NCR nu-nu nude mice. Tumor size was measured every 3 days and a tumor growth curve was created (E) . Points, mean tumor volume; Bars, SEM. (F) , Representative picture of tumor harvested at day 21. (G) , the knockdown of NRP2 expression in tumor samples was confirmed by immunofluorescence staining using NRP2 antibody. Significant differences are indicated by: (*) p < 0.05, (**) p < 0.01. Column: mean value; Error bars: SEM.

Techniques Used: Knockdown, In Vitro, In Vivo, Expressing, shRNA, Real-time Polymerase Chain Reaction, Western Blot, MTT Assay, Soft Agar Assay, Transfection, Control, Plasmid Preparation, Immunofluorescence, Staining

Knockdown of NRP2 inhibited the tumor invasion, migration and lung metastasis of osteosarcoma. (A) , Migration assay. The BD chamber system without Matrigel coating was used to evaluate the migration of shNRP2 and control vector transfected osteosarcoma 143B cells. The migration of 143B cells was significantly inhibited by NRP2 knockdown. (B) , Matrigel invasion assay was performed in BD chamber system coated with Matrigel, using shNRP2 and control vector transfected osteosarcoma 143B cells. There were less NRP2 depleted 143B cells invaded through the matrigel coated porous membrane. (C) , Knockdown of NRP2 in 143B cells reduced the lung metastasis of osteosarcoma in an orthotopic lung metastasis mouse model. Mouse lungs were fixed in Bouin’s solution, and the number of lung surface metastatic nodules was counted and graphed. Each group contained 10 mice and the experiment was repeated 3 times (D) , Representative photographs of lungs with metastatic nodules of osteosarcoma. (E) , NRP2 knockdown significantly reduced both 143B and Saos-2 cells adherence to the endothelial monolayer. The mean cell number was calculated from 10 fields (×100). (F) , Representative images of GFP transfected tumor cells adhering to the endothelial monolayer. Significant differences are indicated by: (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Column: mean value; Error bars: SEM.

Figure Legend Snippet: Knockdown of NRP2 inhibited the tumor invasion, migration and lung metastasis of osteosarcoma. (A) , Migration assay. The BD chamber system without Matrigel coating was used to evaluate the migration of shNRP2 and control vector transfected osteosarcoma 143B cells. The migration of 143B cells was significantly inhibited by NRP2 knockdown. (B) , Matrigel invasion assay was performed in BD chamber system coated with Matrigel, using shNRP2 and control vector transfected osteosarcoma 143B cells. There were less NRP2 depleted 143B cells invaded through the matrigel coated porous membrane. (C) , Knockdown of NRP2 in 143B cells reduced the lung metastasis of osteosarcoma in an orthotopic lung metastasis mouse model. Mouse lungs were fixed in Bouin’s solution, and the number of lung surface metastatic nodules was counted and graphed. Each group contained 10 mice and the experiment was repeated 3 times (D) , Representative photographs of lungs with metastatic nodules of osteosarcoma. (E) , NRP2 knockdown significantly reduced both 143B and Saos-2 cells adherence to the endothelial monolayer. The mean cell number was calculated from 10 fields (×100). (F) , Representative images of GFP transfected tumor cells adhering to the endothelial monolayer. Significant differences are indicated by: (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Column: mean value; Error bars: SEM.

Techniques Used: Knockdown, Migration, Control, Plasmid Preparation, Transfection, Invasion Assay, Membrane

Knockdown of NRP2 resulted in decreased blood vessel density of OS in vivo . (A) , Blood vessels (top panel) and capillaries (bottom panel) in tumor samples were visualized by immunohistochemistry with CD31 antibody. Tumor cell nuclear was stained with DAPI. Number of blood vessels per field (100×) was calculated and graphed (Right). Column, mean number of blood vessel per field (x100); Error bars, SEM. (B & C) , Matrigel tube formation assay: no difference was found in the number of tubules formed by HUVEC in conditioned medium from shNRP2 OS cells and shRNA control cells. (B) Representative photographs of tubules formed by HUVEC cells on Matrigel. (C) The tubular number was calculated and graphed, Column: mean number of tubules; Error bars: SEM. (D) , tumor-endothelial co-culture tube formation assay: The HUVEC cells were stained with CellTracker Red CMTPX dye and the tumor cells were transfected with shNRP2 vector with GFP expression. Left panel: tubules formed by HUVEC (red); middle panel: tumor cells (green) attached on tubules; right panel: merged image (right) of HEVEC tubules (red) and attached tumor cells (green). NRP2 depleted tumor cells sustained distinct morphologic changes compared to control cells (bottom panel).

Figure Legend Snippet: Knockdown of NRP2 resulted in decreased blood vessel density of OS in vivo . (A) , Blood vessels (top panel) and capillaries (bottom panel) in tumor samples were visualized by immunohistochemistry with CD31 antibody. Tumor cell nuclear was stained with DAPI. Number of blood vessels per field (100×) was calculated and graphed (Right). Column, mean number of blood vessel per field (x100); Error bars, SEM. (B & C) , Matrigel tube formation assay: no difference was found in the number of tubules formed by HUVEC in conditioned medium from shNRP2 OS cells and shRNA control cells. (B) Representative photographs of tubules formed by HUVEC cells on Matrigel. (C) The tubular number was calculated and graphed, Column: mean number of tubules; Error bars: SEM. (D) , tumor-endothelial co-culture tube formation assay: The HUVEC cells were stained with CellTracker Red CMTPX dye and the tumor cells were transfected with shNRP2 vector with GFP expression. Left panel: tubules formed by HUVEC (red); middle panel: tumor cells (green) attached on tubules; right panel: merged image (right) of HEVEC tubules (red) and attached tumor cells (green). NRP2 depleted tumor cells sustained distinct morphologic changes compared to control cells (bottom panel).

Techniques Used: Knockdown, In Vivo, Immunohistochemistry, Staining, Tube Formation Assay, shRNA, Control, Co-Culture Assay, Transfection, Plasmid Preparation, Expressing

NRP2 knockdown suppressed endothelial recruitment by osteosarcoma cells. (A) , in a trans-well co-culture migration model, less endothelial cells migrated through the porous insert membrane of the Chamber in shNRP2 group compared to control group. shNRP2 depleted osteosarcoma cells vs control cells were seeded in the lower chamber. HUVECs were seeded in the top insert chamber. Insert picture: Representative photograph of migrated HUVECs. (B) , in a trans-well co-culture invasion model, less endothelial cells invaded through the matrigel coated porous insert membrane of the Chamber in shNRP2 group compared to control group. shNRP2 depleted osteosarcoma cells vs control cells were seeded in the lower chamber. HUVECs were seeded in the top insert chamber. Insert picture: Representative photograph of invaded HUVECs. Significant differences are indicated by: (*) p < 0.05, (**) p < 0.01. Column: mean migrated/invaded cells per field (40×); Error bars: SEM.

Figure Legend Snippet: NRP2 knockdown suppressed endothelial recruitment by osteosarcoma cells. (A) , in a trans-well co-culture migration model, less endothelial cells migrated through the porous insert membrane of the Chamber in shNRP2 group compared to control group. shNRP2 depleted osteosarcoma cells vs control cells were seeded in the lower chamber. HUVECs were seeded in the top insert chamber. Insert picture: Representative photograph of migrated HUVECs. (B) , in a trans-well co-culture invasion model, less endothelial cells invaded through the matrigel coated porous insert membrane of the Chamber in shNRP2 group compared to control group. shNRP2 depleted osteosarcoma cells vs control cells were seeded in the lower chamber. HUVECs were seeded in the top insert chamber. Insert picture: Representative photograph of invaded HUVECs. Significant differences are indicated by: (*) p < 0.05, (**) p < 0.01. Column: mean migrated/invaded cells per field (40×); Error bars: SEM.

Techniques Used: Knockdown, Co-Culture Assay, Migration, Membrane, Control

NRP2 expression is regulated by Wnt signaling pathway. A , Genechip® microarray showed the down-regulated expression of NRP2 in Wnt antagonist sLRP5 transfected osteosarcoma Saos-2 cells. VEGF expression is intact. Real time PCR (B) and western blot (C) with accompanying densitometric assay (D) confirmed the down-regulated mRNA and protein level of NRP2 in Wnt antagonist sLRP5 transfected Saos-2 cells, while VEGF level remains intact. E , western blot and accompanying densitometry showed the down-regulated NRP2 expression in additional Wnt antagonist DKK3 transfected Saos-2 cells, 143B cells and U2-OS cells, and Wif-1 transfected 143B cells. F , Chromatin immunoprecipitation (ChIP) assay verified the binding of TCF4 on the five binding sites in NRP2 promoter region. G , schematic representation of the binding sites of TCF/LEF on the 3-kb promoter region of NRP2 genes.

Figure Legend Snippet: NRP2 expression is regulated by Wnt signaling pathway. A , Genechip® microarray showed the down-regulated expression of NRP2 in Wnt antagonist sLRP5 transfected osteosarcoma Saos-2 cells. VEGF expression is intact. Real time PCR (B) and western blot (C) with accompanying densitometric assay (D) confirmed the down-regulated mRNA and protein level of NRP2 in Wnt antagonist sLRP5 transfected Saos-2 cells, while VEGF level remains intact. E , western blot and accompanying densitometry showed the down-regulated NRP2 expression in additional Wnt antagonist DKK3 transfected Saos-2 cells, 143B cells and U2-OS cells, and Wif-1 transfected 143B cells. F , Chromatin immunoprecipitation (ChIP) assay verified the binding of TCF4 on the five binding sites in NRP2 promoter region. G , schematic representation of the binding sites of TCF/LEF on the 3-kb promoter region of NRP2 genes.

Techniques Used: Expressing, Microarray, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Chromatin Immunoprecipitation, Binding Assay

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