nid1 plasmids  (Addgene inc)

Journal: Therapeutic Advances in Medical Oncology

Article Title: Prostate cancer-associated urinary proteomes differ before and after prostatectomy

doi: 10.1177/17588359221131532

Figure Lengend Snippet: Prediction of the potential tumor suppressors in the urine samples from the post-prostatectomy patients. The single and double asterisks indicate p < 0.05 and 0.01, respectively. (a) List of 41 potential tumor suppressors that were significantly enriched in the urine samples from the post-prostatectomy patients. (b) MTT-based cell viability of TRAMP prostate tumor cells, and 4T1.2 and EO771 mammary tumor cells in response to 19 recombinant proteins at 5 µg/mL. Three proteins (PRSS8, PVRL2, and NID1) were selected for further analysis. (c–e) Concentrations of prostasin (PRSS8), nectin 2 (PVRL2), and NID1 in urine from the pre and post-prostatectomy patients. NID1, nidogen 1.

Article Snippet: The overexpression of NID1 was achieved by transfecting NID1 plasmids (#12016, Addgene, Cambridge, MA, USA).

Techniques: Recombinant

Journal: Therapeutic Advances in Medical Oncology

Article Title: Prostate cancer-associated urinary proteomes differ before and after prostatectomy

doi: 10.1177/17588359221131532

Figure Lengend Snippet: Antitumor effects of PRSS8, PVRL2, and NID1 on TRAMP prostate tumor cells. The single and double asterisks indicate p < 0.05 and 0.01, respectively. (a) Elevated levels of PRSS8, PVRL2, and NID1 in the urine samples from the post-prostatectomy patients. (b and c) Downregulation of TGFβ, and upregulation of c-cas3 and p53 by the incubation of TRAMP prostate tumor cells with 1, 2 µg/mL of PRSS8 and PVRL2. (d and e) Reduction in the EdU-based proliferation and scratch-based migration of TRAMP cells by PRSS8 recombinant proteins. (f and g) Reduction in the EdU-based proliferation and scratch-based migration of TRAMP cells by PVRL2 recombinant proteins. c-cas3, cleaved caspase 3; CN, control; NID1, nidogen 1; PRSS8, prostasin; PVRL2, poliovirus receptor-related 2 or nectin 2; TGFβ, transforming growth factor beta.

Article Snippet: The overexpression of NID1 was achieved by transfecting NID1 plasmids (#12016, Addgene, Cambridge, MA, USA).

Techniques: Incubation, Migration, Recombinant, Control

Journal: Therapeutic Advances in Medical Oncology

Article Title: Prostate cancer-associated urinary proteomes differ before and after prostatectomy

doi: 10.1177/17588359221131532

Figure Lengend Snippet: Differential role of NID1 in the extracellular and intracellular domains for TRAMP prostate tumor cells. The single and double asterisks indicate p < 0.05 and 0.01, respectively. (a) Extracellular NID1 as a tumor suppressor by downregulating TGFβ and upregulating c-cas3 for apoptosis induction and p53 for tumor suppression. (b and c) Reduction in EdU-based proliferation and scratch-based motility of TRAMP cells by extracellular NID1. (d and e) Elevation in the EdU-based proliferation and scratch-based migration of TRAMP cells by the overexpression of NID1. (f) Upregulation of TGFβ and Snail, and downregulation of c-cas3 in TRAMP cells by the overexpression of NID1. (g and h) Downregulation of TGFβ and Snail, and upregulation of c-cas3 in TRAMP cells by the overexpression of PRSS8 and PVRL2. CN, control; c-cas3, cleaved caspase 3; NID1, nidogen 1; TGFβ, transforming growth factor beta.

Article Snippet: The overexpression of NID1 was achieved by transfecting NID1 plasmids (#12016, Addgene, Cambridge, MA, USA).

Techniques: Migration, Over Expression, Control

Journal: Therapeutic Advances in Medical Oncology

Article Title: Prostate cancer-associated urinary proteomes differ before and after prostatectomy

doi: 10.1177/17588359221131532

Figure Lengend Snippet: Tumor selectivity, effects on percent survival, and the putative regulatory mechanism. (a) Tumor selectivity. The tumor selectivity was defined as the ratio of ‘reduction in MTT-based viability of tumor cells’ to ‘reduction in MTT-based viability of non-tumor cells’. The double asterisk indicates p < 0.01. Two tumor cell lines (TRAMP and PC-3 cells) and three non-tumor cell lines (A5, adipose, and MC3T3 cells) were employed. Of note, N.D. means that the viability of non-tumor cells was stimulated. (b–d) Percent survival of all cancer patients with the low and high mRNA levels of PRSS8, PVRL2, and NID1. (e) Putative tumor-suppressing mechanism by urine from the post-prostatectomy patients. NID1, nidogen 1; PSA, prostate-specific antigen.

Article Snippet: The overexpression of NID1 was achieved by transfecting NID1 plasmids (#12016, Addgene, Cambridge, MA, USA).

Techniques:

Journal: Oncology Letters

Article Title: Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

doi: 10.3892/ol.2020.12313

Figure Lengend Snippet: Characteristics of murine mammary tumor cells.

Article Snippet: To reduce Nid1 expression, RJ423 cells were transfected with four unique shRNAs targeting Nid1 (cat. no. TL501474; Origene Technologies Inc.) or a control shRNA (cat. no. TR30021; Origene Technologies Inc.).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

doi: 10.3892/ol.2020.12313

Figure Lengend Snippet: Nid1 levels in cells stably expressing Nid1 shRNA. (A) Expression levels of Nid1 mRNA relative to Hprt in parental RJ423, RJ423con, RJ423 Nid1 A and RJ423sh Nid1 D cells. The bars represent the mean value and the error bars represent the SEM. *Indicates significant difference (P<0.05) from RJ423con cells (n=6). (B) Western blot analysis of NID1 protein from cell lysates and conditioned media from RJ423, RJ423con, RJ423sh Nid1 A and RJ423sh Nid1 D cells. β-actin was used as a loading control for the cell lysates while amido black was used to ensure similar loading for the conditioned media samples. Nid1 , nidogen 1; Hprt , hypoxanthine phosphoribosyltransferase; con, control; sh, short hairpin RNA.

Article Snippet: To reduce Nid1 expression, RJ423 cells were transfected with four unique shRNAs targeting Nid1 (cat. no. TL501474; Origene Technologies Inc.) or a control shRNA (cat. no. TR30021; Origene Technologies Inc.).

Techniques: Stable Transfection, Expressing, shRNA, Western Blot

Journal: Oncology Letters

Article Title: Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

doi: 10.3892/ol.2020.12313

Figure Lengend Snippet: Impact of NID1 conditioned media on cell proliferation and apoptosis. (A) Percentage of PH3-positive cells in parental RJ423, RJ423con, RJ423sh Nid1 A and RJ423sh Nid1 D cells. (B) Percentage of apoptotic cells in RJ423, RJ423con, RJ423 Nid1 A and RJ423 Nid1 D cells. Percentage of PH3-positive cells from (C) RJ423sh Nid1 A and (D) RJ423sh Nid1 D cells treated with conditioned media from RJ423, RJ423con, RJ423sh Nid1 A or RJ423sh Nid1 D cells for 24 h. *P<0.05 vs. RJ423con cells (n=3). Nid1 , nidogen 1; con, control; sh, short hairpin RNA; PH3, phospho-histone H3.

Article Snippet: To reduce Nid1 expression, RJ423 cells were transfected with four unique shRNAs targeting Nid1 (cat. no. TL501474; Origene Technologies Inc.) or a control shRNA (cat. no. TR30021; Origene Technologies Inc.).

Techniques: shRNA

Journal: Oncology Letters

Article Title: Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

doi: 10.3892/ol.2020.12313

Figure Lengend Snippet: Impact of NID1 conditioned media on cell migration. Representative images of toluidine blue stained cells on the bottom of collagen-coated transwell inserts for (A) parental RJ423, (B) RJ423con, (C) RJ423sh Nid1 A and (D) RJ423sh Nid1 D cells (magnification, ×4). (E) Quantification of the percent area occupied by parental RJ423, RJ423con, RJ423sh Nid1 A and RJ423sh Nid1 D cells. Migration and invasion of RJ423sh Nid1 D cells when conditioned media from RJ423, RJ423con, RJ423sh Nid1 A or RJ423sh Nid1 D cells were placed (F and G) in the upper chamber of the well with the cells or (H and I) in the lower chamber. (F and H) Schematic indicating the location of the conditioned media and cells. (G and I) Quantitative data from these experiments. *P<0.05 vs. RJ423con cells (n=3). Nid1, nidogen 1; con, control; sh, short hairpin RNA; CM, conditioned media.

Article Snippet: To reduce Nid1 expression, RJ423 cells were transfected with four unique shRNAs targeting Nid1 (cat. no. TL501474; Origene Technologies Inc.) or a control shRNA (cat. no. TR30021; Origene Technologies Inc.).

Techniques: Migration, Staining, shRNA

Journal: Oncology Letters

Article Title: Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

doi: 10.3892/ol.2020.12313

Figure Lengend Snippet: Expression levels of epithelial and mesenchymal genes following Nid1 knockdown. mRNA expression levels of (A) Cdh1 , (B) Vim , (C) Snai1 , (D) Snai2 , (E) Twist1 , (F) Twist2 , (G) Zeb1 or (H) Zeb2 relative to Hprt. *P<0.05 vs. RJ423con cells (n=5). Nid1 , nidogen 1; Hprt , hypoxanthine phosphoribosyltransferase; con, control; sh, short hairpin RNA.

Article Snippet: To reduce Nid1 expression, RJ423 cells were transfected with four unique shRNAs targeting Nid1 (cat. no. TL501474; Origene Technologies Inc.) or a control shRNA (cat. no. TR30021; Origene Technologies Inc.).

Techniques: Expressing, shRNA