nidogen  (R&D Systems)

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

Journal: International journal of molecular sciences

Article Title: Nidogen 2 Overexpression Promotes Hepatosteatosis and Atherosclerosis.

doi: 10.3390/ijms252312782

Figure Lengend Snippet: Figure 1. Expression of NID2 protein is elevated in human atherosclerotic arteries and murine steatotic livers. (A) Representative western blot images for NID2 and β-tubulin protein expression in human atherosclerotic inner curvature (IC) and non-atherosclerotic descending aorta (DA) vascular tissue. The bar diagram shows mean protein levels expressed as a ratio of NID2 to β-tubulin. (B) Representative Western blot images for NID2 (red arrowhead points to the correct band) and GAPDH in the livers of control diet (CD)- and calorie-matched high-fat diet (HFD, 12 weeks)-fed C57BL/6J mice. The bar diagram represents the mean NID2 protein expression (n = 4). Statistical analyses were performed using a two-tailed unpaired t-test (A,B). Data represent mean ± SEM. * p < 0.05, and **** p < 0.0001.

Article Snippet: The following primary antibodies were used: NID2 (Proteintech, Rosemont, IL, USA #13530-1-AP), total ACC (Cell Signaling Technology, Danvers, MA, USA, #3662S), Int.

Techniques: Expressing, Western Blot, Control, Two Tailed Test

Journal: International journal of molecular sciences

Article Title: Nidogen 2 Overexpression Promotes Hepatosteatosis and Atherosclerosis.

doi: 10.3390/ijms252312782

Figure Lengend Snippet: Figure 2. NID2 overexpression enhances liver and epididymal white adipose tissue mass in male mice. (A) The schematic diagram illustrates the experimental plan. Apoe−/−mice were injected with control (Ctrl) and NID2-AAV intraperitoneally, fed a Western diet for 12 weeks, and analyzed. (B–E) Male control and NID2-AAV-injected Apoe−/−mice were utilized to measure NID2 mRNA levels in various organs by qRT-PCR at least in duplicate. Bar diagrams represent mRNA expression in the liver (B, n = 10), kidney (C, n = 6), epididymal white adipose tissue (EpiWAT, D, n = 7–10), and heart (E, n = 10). Bar diagrams show body weight gain (F), plasma total cholesterol (G), fasting blood glucose (H), whole-body fat/lean mass (I), liver weight (J), adipose tissue weight (K), and spleen weight (L) (n = 5–6). A two-tailed unpaired t-test (C,G–K), two-tailed unpaired Mann–Whitney test (B,D,E,L), and two-way ANOVA followed by Sidak post hoc test for multiple comparisons (F) were utilized for statistical analyses. Data represent mean ± SEM. ns: non-significant. * p < 0.05, *** p < 0.001 and **** p < 0.0001.

Article Snippet: The following primary antibodies were used: NID2 (Proteintech, Rosemont, IL, USA #13530-1-AP), total ACC (Cell Signaling Technology, Danvers, MA, USA, #3662S), Int.

Techniques: Over Expression, Injection, Control, Western Blot, Quantitative RT-PCR, Expressing, Clinical Proteomics, Two Tailed Test, MANN-WHITNEY

Journal: International journal of molecular sciences

Article Title: Nidogen 2 Overexpression Promotes Hepatosteatosis and Atherosclerosis.

doi: 10.3390/ijms252312782

Figure Lengend Snippet: Figure 3. NID2 overexpression in mice promotes hepatic lipid accumulation and fibrosis. Male Apoe−/−mice were injected with control and NID2-AAV intraperitoneally, fed a Western diet for 12 weeks, and analyzed. (A) Representative Western blot images for NID2 and GAPDH protein expression in the livers of control and NID2-overexpressing mice (n = 3). (B) Representative images of liver sections stained with H & E (lipid droplets), ORO (neutral lipid accumulation), and Sirius red (fibrosis); scale bar 100 µm. (C–H) Bar diagrams represent lipid accumulation (C, n = 6), fibrosis area (D, n = 5), hepatic triglyceride (E, n = 3–4), NEFA levels (F, n = 3–4), plasma triglyceride (G, n = 5) and NEFA levels (H, n = 5), in control and NID2-AAV-injected mice. Statistical analyses were performed using a two-tailed unpaired t-test (C–H). Data represent mean ± SEM. ns: non-significant. * p < 0.05, and ** p < 0.01.

Article Snippet: The following primary antibodies were used: NID2 (Proteintech, Rosemont, IL, USA #13530-1-AP), total ACC (Cell Signaling Technology, Danvers, MA, USA, #3662S), Int.

Techniques: Over Expression, Injection, Control, Western Blot, Expressing, Staining, Clinical Proteomics, Two Tailed Test

Journal: International journal of molecular sciences

Article Title: Nidogen 2 Overexpression Promotes Hepatosteatosis and Atherosclerosis.

doi: 10.3390/ijms252312782

Figure Lengend Snippet: Figure 4. NID2 overexpression augments atherosclerosis in male hypercholesterolemic mice. Male Apoe−/−mice were injected with control (Ctrl) and NID2-AAV intraperitoneally, fed a Western diet for 12 weeks and analyzed. (A) Representative in situ images of the aortic arch (red arrowheads point to atherosclerotic lesions). (B) Representative ORO staining of whole aortas; scale bar 5 mm. The bar diagram represents ORO-positive areas in whole aortas (n = 6). (C) Representative images of aortic root cross-sections stained with H & E (lesion area and necrotic core), ORO (lipid accumulation), and Masson’s trichrome (collagen content); scale bar 200 µm. (D–G) Bar diagrams show lesion area (D), lipid deposition (E), collagen content (F), and necrotic core area (G) (n = 5–6). Statistical analyses were performed using a two-tailed unpaired t-test (B,D–G). Data represent mean ± SEM. * p < 0.05, and ** p < 0.01.

Article Snippet: The following primary antibodies were used: NID2 (Proteintech, Rosemont, IL, USA #13530-1-AP), total ACC (Cell Signaling Technology, Danvers, MA, USA, #3662S), Int.

Techniques: Over Expression, Injection, Control, Western Blot, In Situ, Staining, Two Tailed Test

Journal: International journal of molecular sciences

Article Title: Nidogen 2 Overexpression Promotes Hepatosteatosis and Atherosclerosis.

doi: 10.3390/ijms252312782

Figure Lengend Snippet: Figure 5. NID2 overexpression inhibits the activation of the lipid metabolism-related protein AMPK. (A) Representative Western blot images for lipid metabolism and pro-inflammatory proteins utilizing liver lysates from control and NID2-AAV-injected mice. Bar diagrams represent mean protein expression (B,C) as the ratios of phospho-total proteins ACC (B) and AMPK (C), and protein levels of IL-6 (D) and TNFα (E) (n = 5). Statistical analyses were performed using a two-tailed unpaired t-test. Data represent mean ± SEM. ns: non-significant. * p < 0.05.

Article Snippet: The following primary antibodies were used: NID2 (Proteintech, Rosemont, IL, USA #13530-1-AP), total ACC (Cell Signaling Technology, Danvers, MA, USA, #3662S), Int.

Techniques: Over Expression, Activation Assay, Western Blot, Control, Injection, Expressing, Two Tailed Test