Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: The common DEGs of four gene expression profiles (adj. P value <0.05, |logFC|> 2.0).

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques: Gene Expression

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: PPI network and hub genes' identification. (a) PPI network was structured by the 54 upregulated DEGs using STRING database. (b) The top 10 hub genes in the PPI network were screened ground on their connectivity degree. The genes such as CD44, COL1A1, COL1A2, COL3A1, FN1, IGFBP3, LOX, POSTN, TIMP1, and VEGFA are represented from red (high degree) to yellow (low degree). (c) Gene Ontology (GO) chord diagram of the top genes in GBM.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques:

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: Top ten hub genes with higher degree of connectivity.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques: Binding Assay

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: The AUC analysis of hub genes. AUC analysis of CD44, COL1A1, COL1A2, COL3A1, FN1, IGFBP3, LOX, POSTN, TIMP1, and VEGFA for distinguishing GBM samples from normal tissues.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques:

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: OS, DFS, and univariate and multivariate Cox analysis of the hub genes overexpressed in GBM patients. (a) OS of the hub genes overexpressed in GBM sufferers. (b) DFS of the hub genes overexpressed in GBM sufferers. (c) Univariate and multivariate Cox analysis of TIMP1 expression and other clinical pathological factors for OS. HR, hazard ratio.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: Correlation between immune infiltrates and TIMP1 expression in GBM. (a) TIMP1 expression was definitely connected with CD4+ T cell, B cell, T cell regulatory (Tregs), neutrophils, cancer-associated fibroblast, macrophage M1, and myeloid dendritic cell. (b–d) Higher infiltration of Tregs, cancer-associated fibroblast, and mast cell correlated with worse prognosis.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: Correlation analysis between TIMP1 and related genes and markers of immune cells in TIMER.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques:

Journal: Journal of Oncology

Article Title: Identification of Prognostic Biomarkers of Glioblastoma Based on Multidatabase Integration and Its Correlation with Immune-Infiltration Cells

doi: 10.1155/2022/3909030

Figure Lengend Snippet: Downregulation of TIMP1 inhibits cell proliferation, migration, and invasion in U-87 MG and U-118 MG cells. (a, b) The results of qPCR and WB showed that compared with many glioblastoma cell lines, U-87 MG and U-118 MG cell lines were selected for TIMP1 gene related function test. (c, d) qPCR and WB showed that TIMP1 expression was inhibited in U-87 MG and U-118 MG cells transfected with sh-TIMP1 compared with the NC group. (e, f) CCK-8 and colony formation assays revealed that the proliferation abilities were suppressed in U-87 MG and U-118 MG cells transfected with sh-TIMP1 compared with the NC group. (g, h) Transwell assay revealed that cell migration and invasion were suppressed in U-87 MG and U-118 MG cells transfected with sh-TIMP1 compared with the NC group. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: After blocking with 5% fat-free milk, incubate with primary antibodies against TIMP1 (A00561-1; Boster, China) and β -actin (Beyotime Biotechnology, China), respectively, using a dilution of 1 : 1000 for each antibody overnight at 4°C.

Techniques: Migration, Expressing, Transfection, CCK-8 Assay, Transwell Assay

Journal: Scientific Reports

Article Title: Cellular uptake of proMMP-2:TIMP-2 complexes by the endocytic receptor megalin/LRP-2

doi: 10.1038/s41598-017-04648-y

Figure Lengend Snippet: Time-course of cell-surface binding of 125 I-proMMP-2:TIMP-2 complex. BN16 cells were incubated with 10 nM 125 I-proMMP-2:TIMP-2 complex at 4 °C for the indicated intervals to allow surface binding, in the absence (open circles) or presence of 1 μM RAP (closed circles), then washed and treated with pronase ® for detachment. Surface-bound 125 I-proMMP-2:TIMP-2 complex was defined as pronase ® -sensitive radioactivity. Values are means ± S.D. of three dishes. This experiment was performed twice with similar results.

Article Snippet: The proMMP-2:TIMP-2 complex was purchased from Enzo Life Sciences (Brussels, Belgium).

Techniques: Binding Assay, Incubation, Radioactivity

Journal: Scientific Reports

Article Title: Cellular uptake of proMMP-2:TIMP-2 complexes by the endocytic receptor megalin/LRP-2

doi: 10.1038/s41598-017-04648-y

Figure Lengend Snippet: Blocking of megalin/LRP-2 but not LRP-1 inhibits membrane binding of proMMP-2:TIMP-2 complex. BN16 cells were incubated with 10 nM 125 I-proMMP-2:TIMP-2 complex at 4 °C for 2 h to allow surface binding, in the absence or presence of 1 μM RAP, anti-megalin/LRP-2 IgG (100 μg/ml), anti-LRP-1 IgG (100 μg/ml) or non-immune IgG (100 μg/ml). After 2 h, surface-bound tracer was measured as in Fig. . Values are means ± S.D. of three dishes. **p < 0.001; NS, not significant, vs control, using Student’s t test.

Article Snippet: The proMMP-2:TIMP-2 complex was purchased from Enzo Life Sciences (Brussels, Belgium).

Techniques: Blocking Assay, Binding Assay, Incubation

Journal: Scientific Reports

Article Title: Cellular uptake of proMMP-2:TIMP-2 complexes by the endocytic receptor megalin/LRP-2

doi: 10.1038/s41598-017-04648-y

Figure Lengend Snippet: Uptake and degradation of 125 I- proMMP-2:TIMP-2 complex by rat yolk sac BN16 cells. BN16 cells were incubated with 10 nM 125 I-proMMP-2:TIMP-2 complex at 4 °C for 2 h to allow surface binding, in the absence (open circles) or presence of 1 μM RAP (closed circles). Some of the cultures were pre-treated with 100 μM chloroquine for 1 h at 37 °C (open triangles). After washing, cells were further incubated with fresh medium pre-warmed at 37 °C, without or with RAP or chloroquine as above. At the indicated times, the amounts of ligand ( a ) remaining surface-bound (pronase ® -sensitive radioactivity), ( b ) internalized (pronase ® -resistant radioactivity) and ( c ) degraded (trichloroacetic acid-soluble radioactivity in conditioned medium) were measured. Values are means ± S.D. of three dishes. This experiment was performed twice with similar results.

Article Snippet: The proMMP-2:TIMP-2 complex was purchased from Enzo Life Sciences (Brussels, Belgium).

Techniques: Incubation, Binding Assay, Radioactivity

Journal: Scientific Reports

Article Title: Cellular uptake of proMMP-2:TIMP-2 complexes by the endocytic receptor megalin/LRP-2

doi: 10.1038/s41598-017-04648-y

Figure Lengend Snippet: Invalidation of megalin/LRP-2 induces accumulation of proMMP-2 and TIMP-2 in urines. Twenty-four-hour urines were collected from 7-month-old Meg lox/lox ;Wnt4-Cre + mice (cKO) or their wild-type littermates (WT) and samples were analyzed, without concentration, by direct (32 µl/lane; top panel) or reverse (16 µl/lane; bottom panel) zymography. Reference (ref.) corresponds to medium conditioned by mouse calvarium for direct zymography (as in Fig. ) or 20 ng recombinant TIMP-2 for reverse zymography.

Article Snippet: The proMMP-2:TIMP-2 complex was purchased from Enzo Life Sciences (Brussels, Belgium).

Techniques: Concentration Assay, Zymography, Recombinant