anti nid2 antibody  (Santa Cruz Biotechnology)

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: Extracellular vesicles from metastatic colorectal cancer cells (LM3) promote CRC tumor metastasis (A) Exosomes in the cell supernatants were identified by western blot experiments. (B) Exos in the cell supernatants were identified by transmission electron microscopy (upper) and NanoFCM (bottom). (C) Schematic diagram of the EV education mouse model. Nude mice were injected with EVs derived from SW480-P, SW480-LM3, km12-P, or km12-LM3 cells via the tail vein. Nude mice were injected with EVs derived from SW480-P, SW480-LM3, km12-P, or km12-LM3 cells via the tail vein once a week for 3 weeks (15 μg per week) prior to splenic injection of tumor seeds derived from SW480-P or km12-P cells ( n = 5). Analysis of liver metastases was performed 2 weeks after splenic injection. (D) In vivo fluorescence imaging and quantitative analysis of animals at the end of the experiment. (E) Venn diagram showing the overlap of upregulated genes identified via mRNA-seq in highly metastatic cells, serum exosome proteins in highly metastatic cells, and serum Exo proteins in colorectal cancer patients. (F) Representative IHC staining of NID1 expression in normal colon epithelium and paired primary CRC tissues. Scale bar: 50 μm. (G) NID1 expression was higher in CRC tissues ( n = 174) than in normal colon epithelium tissues ( n = 174) and higher in LM (+) CRC tissues ( n = 15) than in LM (−) CRC tissues ( n = 15) and in LM (−) CRC tissues ( n = 159). (H) Kaplan-Meier survival curves of overall survival in CRC patients with low ( n = 72) or high ( n = 102) NID1 expression. n.s. represents not significant; ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with paired t tests (D and G). Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Western Blot, Transmission Assay, Electron Microscopy, Injection, Derivative Assay, In Vivo, Fluorescence, Imaging, Immunohistochemistry, Expressing

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: The exosomal protein NID1 promotes liver metastasis of CRC tumors (A) EV mouse model comparing the effects of EVs from SW480/km12-LM3 and shNID1-1 SW480/km12-LM3 cells on CRC metastasis ( n = 5). (B and C) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (D) Comparison of the effects of EVs from XPack and XP-NID1 HCT116 cells on CRC metastasis in an EV mouse model ( n = 5). (E and F) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (G) Schematic diagram of the mouse model of in situ cecum injection combined with splenic injection. (H and I) In vivo imaging images of the mice and quantitative fluorescence statistics of the mouse liver region. n.s. represents not significant; ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with a t test (B, C, E, F, H, and I). Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Luciferase, Comparison, In Situ, Injection, In Vivo Imaging, Fluorescence

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: NID1 promotes colorectal cancer metastasis through the exosomal pathway (A–D) Transwell assays were performed to verify the effects of exosome-derived NID1 and the endogenous expression of NID1 on the invasive and metastatic ability of colorectal cancer cells in vitro . (E) Schematic diagram of a mouse spleen injected with tumor cells. (F) Splenic injection of SW480/km12 and NID1-overexpressing SW480/km12 cells in the presence/absence of cilengitide in vivo and NID1-overexpressing SW480/km12 cells overexpressing NID1 in vitro . (G) Quantification of the luciferase signal is shown. n.s. represents not significant; ∗∗∗ represents p < 0.001; ∗∗ represents p < 0.01, analyzed with paired t tests (A–D and G). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Derivative Assay, Expressing, In Vitro, Injection, In Vivo, Luciferase

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: The exosomal protein NID1 promotes epithelial mesenchymal transition in colorectal cancer cells (A) Western blotting was used to verify the effects of the EMT marker exosome-derived NID1 on colorectal cancer cells in vitro . (B) RT-qPCR was used to verify the effect of the EMT marker exosome-derived NID1 on colorectal cancer cells in vitro . (C) Western blotting was used to validate the expression of the EMT marker NID1 endogenously expressed in colorectal cancer cells in vitro . (D) RT-qPCR was used to verify the effect of the endogenous expression of the EMT marker NID1 on colorectal cancer cells in vitro . (E) Correlation of the expression of NID1 and EMT-associated mRNAs in primary tumors from the TCGA CRC patient cohort. (F) Relative mRNA expression (TPM) of EMT markers in LV5 and LV5-NID1 SW480 cells treated with GW4869 (10 μg mL-1) for 72 h. The TPM expression of EMT markers in LV5 and LV5-NID1 SW480 cells is shown in the following table. (G) KEGG pathway analysis revealed enrichment of genes associated with metastatic CRC cell lines. (H) Western blotting was used to verify the effect of the PI3K-Akt pathway marker exosome-derived NID1 on colorectal cancer cells in vitro . n.s. represents not significant; ∗∗ represents p < 0.01, analyzed with paired t test (B, D, and F). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Western Blot, Marker, Derivative Assay, In Vitro, Quantitative RT-PCR, Expressing

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: Ev-NID1 induces NET formation by facilitating IL-11 production in HSCs (A) Representative confocal microscope images showing the uptake of PKH26-labeled exosomes by LX-2 cells. Scale bar: 200 μm. (B) Bubble plot showing the GO signatures enriched in ev-NID1-stimulated LX-2 cells. (C) Immunofluorescence (IF) staining and quantification of NETs (H3Cit+ and MPO+ structures) in the liver metastases of mice treated with exosomes derived from ev-X-Pack HCT-116 and ev-XP-NID1 HCT116 cells for 12 h ( n = 18 Region of Metastatic Focus (RMFs) from 6 experimental replicates per group). Scale bar: 50 μm. (D) Cytokine array of the media of LX-2 cells pretreated with exosomes (X-Pack and XP-NID1) for 24 h. (E) ELISA results showing that IL-11 was upregulated in the supernatant of LX-2 cells stimulated with exosomes from XP-NID1. (F) Western blotting was used to verify the effect of the PI3K-Akt pathway marker exosome-derived NID1 on colorectal cancer cells in vitro . n.s. represents not significant; ∗∗∗ represents p < 0.001, analyzed with a t test (C and E). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Microscopy, Labeling, Immunofluorescence, Staining, Derivative Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Marker, In Vitro

Journal: iScience

Article Title: Nidogen 1-enriched extracellular vesicles promote liver metastasis by inducing EMT and activating stellate cells

doi: 10.1016/j.isci.2025.113975

Figure Lengend Snippet: In liver metastases, NET formation is associated with IL-11, and targeting the IL-11 signaling pathway with a monoclonal antibody prevents CRLM (A) Immunofluorescence (IF) staining and quantification of IL-11+HSCs (IL-11+ and Desmin+) formed by LX-2 cells treated with exosomes derived from ev-X-Pack HCT-116 and ev-XP- NID1 HCT116 cells for 3 weeks. (B) Western blot analysis to characterize the expression of IL-11 in EV-NID1-treated LX-2 cells. (C) Representative confocal microscope images showing IL-11 expression in LX-2 cells treated with exosomes. Scale bar: 200 μm. (D) Western blot analysis of IL-11 expression in LX-2 cells under normal and quiescent conditions after treatment with EV-NID1 and the PI3K inhibitor (LY294002). (E) qPCR analysis of IL-11 expression in LX-2 cells under normal and quiescent conditions following treatment with EV-NID1 and the PI3K inhibitor (LY294002). (F) Splenic injection of XP-SW480/KM12 and XP-NID1 SW480/KM12 cells for liver metastasis experiments in the presence/absence of anti-IL-11. (G) Image showing the luciferase signal of the animals at the end of the experiment. Quantification of the luciferase signal is shown. (H) IF staining and quantification of NETs (H3Cit+ and MPO+ structures) in the liver metastases of mice treated with exosomes derived from ev-X-Pack HCT-116 and ev-XP-NID1 HCT116 cells for 12 h ( n = 18 RMFs from 6 experimental replicates per group). Scale bar: 50 μm. ∗∗ represents p < 0.01; ∗∗∗ represents p < 0.001, analyzed with a t test (A, E, and F). Three independent experiments were performed. Data are represented as mean ± SEM.

Article Snippet: NID1 , Proteintech , Cat. No. 13766-1-AP.

Techniques: Immunofluorescence, Staining, Derivative Assay, Western Blot, Expressing, Microscopy, Injection, Luciferase

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 1. NID2 was upregulated in various cancer types. (A) PCA plot of the 7357 genes filtered by WGCNA in the GSE16011 dataset. (B) The volcano plots of 275 DEGs in GSE16011. (C) The expression values of NID2 for gliomas and the controls were compared using the Wilcoxon Rank-Sum test in GSE16011. (D) NID2 expression values in various cancers and adjacent non-cancerous tissue from the TCGA database. The red rectangles represent NID2 expression in tumor tissues, while the green rectangles represent NID2 expression in the corresponding non-cancerous tissues. (E) NID2 expression in TCGA GBM samples compared with GTEx brain tissue controls. (F,G) NID2 expression in GSE7696 and GSE4290 GBM samples compared with normal controls. The red dots represent NID2 expression in GBM, and the green dots represent NID2 expression in the control brain tissues. *** p < 0.001, ** p < 0.01. Error bars show the standard error. Dim1/2, Dimensionality 1/2.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Expressing, Control

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 2. Tumor subtypes and clinical outcomes associated with NID2 expression in glioma dataset. (A,D) Wilcoxon rank-sum test was used to analyze the differential expression of NID2 between GBM and LGG in TCGA (A) and CGGA (D). (B,E) Violin plot illustrating NID2 expression in TCGA (B) and CGGA (E) dataset according to the grade. (C,F) Kaplan–Meier survival curves of the TCGA (C) and CGGA (F) cohort showed that a high level of NID2 expression was associated with significantly worse overall glioma survival. WHO II, III and IV, World Health Organization grades II, III, and IV. *** p < 0.001.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative Proteomics

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 3. Tumor subtypes and clinical outcomes associated with NID2 expression in CGGA glioma dataset. (A,B) Univariate (A) and multivariate Cox regression (B) analysis demonstrated NID2 as an independent OS factor in CGGA.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 4. Strong immunoreactivity of NID2 in glioma pathology specimens correlates with high tumor grade. (A–D). Representative photomicrographs of NID2 IHC staining patterns for normal brain tissue (negative, (A)), grade II glioma (mild, (B)), grade III glioma (moderate, (C)), and GBM (strong, (D)) as visualized in 4× (left panel) and 40× (right panel) magnifications. (E) Average NID2 immunoreactive score of LGG versus GBM with corresponding 95% confidence interval error bars. The Wilcox test demonstrated a significant difference between the NID2 immunoreactive score in LGGs and GBMs (p < 0.001). (F) Heatmap of NID2 immunoreactive score distribution according to tumor grade, type, clinical characteristics, and PD-L1 expression in TMA glioma samples. WHO II, III and IV, World Health Organization grades II, III, and IV. *** p < 0.001.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Immunohistochemistry, Expressing

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 5. NID2 regulates the proliferation and migration of glioma cells. (A) Western blotting results of NID2 expression in the vector controls and NID2-overexpressing U87MG/T98G glioma cells. (B) RT-qPCR results of NID2 expression in the vector controls and NID2-overexpressing U87MG/T98G glioma cells. (C) Venn diagram represents genes upregulated in T98G and U87MG glioma cells. (D) The volcano plot of DEGs was between the vector controls and the NID2- overexpressing cells. (E) Gene ontology enrichment analysis of DEGs regulated by NID2 over- expression. GO enrichment analysis contains biological process (BP) and cellular component (CC). DEGs, differentially expressed genes. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Migration, Western Blot, Expressing, Plasmid Preparation, Quantitative RT-PCR, Over Expression

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 6. Overexpression of NID2 promoted the proliferation of glioma cells. (A,B) CCK-8 assay showed that the upregulation of NID2 expression in glioma cells resulted in increased cell pro- liferation (n = 8) in T98G (A) and U87MG (B) glioma cells. (C,D) The represented image of the EdU assay showed that NID2 overexpression in T98G and U87MG cells promoted cell proliferation (Magnification: 10×, n = 3). (E,F) Histograms represent the percentage of the EdU-positive cells. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Over Expression, CCK-8 Assay, Expressing, EdU Assay

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 7. Overexpression of NID2 promoted migration and invasion of glioma cells. (A,B) Wound healing assay showed overexpression of NID2 enhanced glioma cell migration (representative images of wound scratch). (C,D) Overexpression of NID2 promoted the migration and invasion of glioma cells examined by transwell assay. (E–G) Histograms represent the analysis of the wound healing rate (E), migration cell number (F), and invasion cell number (G). *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Over Expression, Migration, Wound Healing Assay, Transwell Assay

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 8. Overexpression of NID2 protected against glioma cell apoptosis. (A) KEGG enrichment analysis of the DEGs between the vector control groups and NID2-overexpressing T98G/U87MG glioma cells revealed significant activation of Akt signaling and extracellular matrix remodeling pathways. (B) GSEA demonstrated that NID2-overexpressing glioma cells exhibited enhanced negative regulation of apoptosis and Akt pathway activation. (C) The TUNEL assay revealed a significant decrease in apoptotic cells in the NID2-overexpressing cells compared to the vector controls (Scale bar: 20 µm). (D,E) Caspase 8 and caspase 3/7 activity assays showed the apoptosis proteins were downregulated in the NID2 overexpression group (n = 8). (F) Western blotting of apoptosis markers in T98G glioma cells. **** p < 0. 0001, *** p < 0.001, ** p < 0.01.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Over Expression, Plasmid Preparation, Control, Activation Assay, TUNEL Assay, Activity Assay, Western Blot

Journal: International journal of molecular sciences

Article Title: NID2 Affects Prognosis of Glioma via Activating the Akt Signaling Pathway.

doi: 10.3390/ijms26083859

Figure Lengend Snippet: Figure 9. Blockage of Akt signaling dampened the anti-apoptic effect of NID2 overexpression. (A) Comparison of apoptotic cells visualized by the TUNEL assay. (B) The activation of Bcl-xL anti-apoptic protein by NID2 overexpression could be reversed by Akt inhibition. **** p < 0.0001, ** p < 0.01, * p < 0.05.

Article Snippet: NID2 was detected using an anti-NID2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-373859) at 1:500 dilution according to the manufacturer’s instructions.

Techniques: Over Expression, Comparison, TUNEL Assay, Activation Assay, Inhibition