Journal: Open Access Rheumatology : Research and Reviews

Article Title: A suppressive effect of prostaglandin E 2 on the expression of SERPINE1 /plasminogen activator inhibitor-1 in human articular chondrocytes: An in vitro pilot study

doi:

Figure Lengend Snippet: Suppressive effect of PGE 2 on PAI-1 expression. A ) Western blot. Left: Chondrocytes were stimulated with PGE 2 (0.1, 1, or 10 nM) for 24 h, and the expression of PAI-1 was analyzed. NS: not stimulated. Representative results are shown. Bottom: Calculation of the relative intensity of the bands among four OA samples tested. B ) ELISA. Levels of PAI-1 in culture supernatants of chondrocytes after PGE 2 stimulation (0.1–10 nM) were measured. Notes: OA chondrocytes, N = 3, duplicate. Abbreviations: ELISA, enzyme-linked immunosorbent assay; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; PAI-1, plasminogen activator inhibitor; PGE 2 , prostaglandin E 2 .

Article Snippet: The level of PAI-1 in the culture supernatant was measured by using an ELISA kit (AssayMax Human Plasminogen activator inhibitor-1 ELISA Kit™, AssayPro LLC, St. Charles, MO, USA).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Open Access Rheumatology : Research and Reviews

Article Title: A suppressive effect of prostaglandin E 2 on the expression of SERPINE1 /plasminogen activator inhibitor-1 in human articular chondrocytes: An in vitro pilot study

doi:

Figure Lengend Snippet: The PAI-1 suppression by PGE 2 is delivered through EP4 receptor. The summary of PAI-1 ELISA with receptor antagonists are shown. Chondrocytes were stimulated with PGE 2 (10 nM) with or without AH6809 (10 ng/ml) or GW627368X (5 μM), and the levels of PAI-1 in culture supernatants were measured. Notes: OA chondrocytes, N = 6. Abbreviations: ELISA, enzyme-linked immunosorbent assay; PAI-1, plasminogen activator inhibitor; PGE 2 , prostaglandin E 2 .

Article Snippet: The level of PAI-1 in the culture supernatant was measured by using an ELISA kit (AssayMax Human Plasminogen activator inhibitor-1 ELISA Kit™, AssayPro LLC, St. Charles, MO, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Open Access Rheumatology : Research and Reviews

Article Title: A suppressive effect of prostaglandin E 2 on the expression of SERPINE1 /plasminogen activator inhibitor-1 in human articular chondrocytes: An in vitro pilot study

doi:

Figure Lengend Snippet: Inhibition of p38 and ERK MAPK did not abrogate the effect of PGE 2 on PAI-1 downregulation. The summary of PAI-1 ELISA with receptor antagonists are shown. Chondrocytes were stimulated with PGE 2 (10 nM) with or without SB203580 or PD98059, and the levels of PAI-1 in culture supernatants were measured. Notes: OA chondrocytes, N = 3. *p <0.05; **p <0.02. Abbreviations: MAPK, mitogen-activated protein kinases, PAI-1, plasminogen activator inhibitor; PGE 2 , prostaglandin E 2 .

Article Snippet: The level of PAI-1 in the culture supernatant was measured by using an ELISA kit (AssayMax Human Plasminogen activator inhibitor-1 ELISA Kit™, AssayPro LLC, St. Charles, MO, USA).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: World Journal of Gastrointestinal Oncology

Article Title: MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis

doi: 10.4251/wjgo.v16.i5.2123

Figure Lengend Snippet: Primer sequences used in this study

Article Snippet: The proteins were then boiled at 100°C for 10 min with 100 μL of loading buffer (Beyotime Biotechnology, China) and separated by SDS-PAGE at 120 V. After electrophoresis, the proteins were transferred onto PVDF membranes and blocked using 5% BSA/TBST for 60 min. the primary rabbit polyclonal antibodies, SERPINE1 (Proteintech, United States), α-smooth muscle actin (α-SMA) (Proteintech, United States), β-catenin (Proteintech, United States), vimentin (HuaBio, China), ERK1/2 (HuaBio, China), phosphorylation extracellular signal-regulated kinase1/2 (p-ERK1/2) (HuaBio, China) and actin (ZSGB-BIO, China), and the mouse monoclonal antibody E-cad (HuaBio, China), were added to the membranes and incubated overnight at 4 °C.

Techniques:

Journal: World Journal of Gastrointestinal Oncology

Article Title: MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis

doi: 10.4251/wjgo.v16.i5.2123

Figure Lengend Snippet: Serpin family E member 1 is a target of microRNA-145-5p. A: miRNA-145-5p and serpin family E member 1 ( SERPINE1 ) 3'- UTR binding sites were predicted using TargetScan; B: WT-SERPINE1 and MUT-SERPINE1 sequences; C: Relative luciferase activity of 293T cells transfected with either WT-SERPINE1 or MUT-SERPINE1 . c P < 0.001.

Article Snippet: The proteins were then boiled at 100°C for 10 min with 100 μL of loading buffer (Beyotime Biotechnology, China) and separated by SDS-PAGE at 120 V. After electrophoresis, the proteins were transferred onto PVDF membranes and blocked using 5% BSA/TBST for 60 min. the primary rabbit polyclonal antibodies, SERPINE1 (Proteintech, United States), α-smooth muscle actin (α-SMA) (Proteintech, United States), β-catenin (Proteintech, United States), vimentin (HuaBio, China), ERK1/2 (HuaBio, China), phosphorylation extracellular signal-regulated kinase1/2 (p-ERK1/2) (HuaBio, China) and actin (ZSGB-BIO, China), and the mouse monoclonal antibody E-cad (HuaBio, China), were added to the membranes and incubated overnight at 4 °C.

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection

Journal: World Journal of Gastrointestinal Oncology

Article Title: MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis

doi: 10.4251/wjgo.v16.i5.2123

Figure Lengend Snippet: Serpin family E member 1 is a potential target gene of microRNA-145-5p and is highly expressed in gastric cancer tissues and cells. A: A Venn diagram of overlapping mRNAs from the differentially expressed mRNAs in The Cancer Genome Atlas (TCGA)-STAD data and miRNA-145-5p ( miR-145-5p ) target genes predicted from the two bioinformatics databases; B: Box diagram showing the differential expression of serpin family E member 1 ( SERPINE1 ) in the normal and tumor data in TCGA-STAD; C: Overall survival curves of patients in the high (red) and low (blue) SERPINE1 expression groups; D: Differential expression of SERPINE1 mRNA (left panel) and protein (middle panel) in gastric cancer (GC) tissue and SERPINE1 mRNA (right panel) in the HGC27 and SGC7901 GC cell lines; scale bar = 20 μm; E: SERPINE1 mRNA and protein expression in the miR-145-5p mimic group in SGC7901 cells and the inhibitor group in HGC27 cells. b P < 0.01; c P < 0.001; d P < 0.0001.

Article Snippet: The proteins were then boiled at 100°C for 10 min with 100 μL of loading buffer (Beyotime Biotechnology, China) and separated by SDS-PAGE at 120 V. After electrophoresis, the proteins were transferred onto PVDF membranes and blocked using 5% BSA/TBST for 60 min. the primary rabbit polyclonal antibodies, SERPINE1 (Proteintech, United States), α-smooth muscle actin (α-SMA) (Proteintech, United States), β-catenin (Proteintech, United States), vimentin (HuaBio, China), ERK1/2 (HuaBio, China), phosphorylation extracellular signal-regulated kinase1/2 (p-ERK1/2) (HuaBio, China) and actin (ZSGB-BIO, China), and the mouse monoclonal antibody E-cad (HuaBio, China), were added to the membranes and incubated overnight at 4 °C.

Techniques: Quantitative Proteomics, Expressing

Journal: World Journal of Gastrointestinal Oncology

Article Title: MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis

doi: 10.4251/wjgo.v16.i5.2123

Figure Lengend Snippet: Effects of serpin family E member 1 overexpression on gastric cancer cell proliferation, invasion, migration, and apoptosis. A: Serpin family E member 1 ( SERPINE1 ) mRNA and protein expression in lentivirus transfected SGC7901 cells; B: SERPINE1 overexpression promotes SGC7901 cell proliferation; C: SERPINE1 overexpression promotes SGC7901 cell colony formation; D: SERPINE1 promotes SGC7901 cell invasion; scale bar = 100 μm; E: SERPINE1 promotes SGC7901 cell migration; scale bar = 100 μm; F: SERPINE1 overexpression reduces SGC7901 cell apoposis. a P < 0.05; b P < 0.01; c P < 0.001.

Article Snippet: The proteins were then boiled at 100°C for 10 min with 100 μL of loading buffer (Beyotime Biotechnology, China) and separated by SDS-PAGE at 120 V. After electrophoresis, the proteins were transferred onto PVDF membranes and blocked using 5% BSA/TBST for 60 min. the primary rabbit polyclonal antibodies, SERPINE1 (Proteintech, United States), α-smooth muscle actin (α-SMA) (Proteintech, United States), β-catenin (Proteintech, United States), vimentin (HuaBio, China), ERK1/2 (HuaBio, China), phosphorylation extracellular signal-regulated kinase1/2 (p-ERK1/2) (HuaBio, China) and actin (ZSGB-BIO, China), and the mouse monoclonal antibody E-cad (HuaBio, China), were added to the membranes and incubated overnight at 4 °C.

Techniques: Over Expression, Migration, Expressing, Transfection

Journal: World Journal of Gastrointestinal Oncology

Article Title: MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis

doi: 10.4251/wjgo.v16.i5.2123

Figure Lengend Snippet: Serpin family E member 1 reverses microRNA-145-5p-mediated gastric cancer cell proliferation, invasion, metastasis, epithelial-mesenchymal transition, and apoptosis. A: Serpin family E member 1 protein expression in co-transfected cells; B: Co-transfected cell proliferation; C: Co-transfected cell colony formation; D: Co-transfected cell migration; scale bar = 100 μm; E: Co-transfected cell invasion; scale bar = 100 μm; F: Epithelial-mesenchymal transition-related marker protein (E-cadherin, β-catenin, Vimentin, and α-smooth muscle actin) expression in co-transfected cells; G: Apoptosis of co-transfected cells. a P < 0.05; b P < 0.01; c P < 0.001.

Article Snippet: The proteins were then boiled at 100°C for 10 min with 100 μL of loading buffer (Beyotime Biotechnology, China) and separated by SDS-PAGE at 120 V. After electrophoresis, the proteins were transferred onto PVDF membranes and blocked using 5% BSA/TBST for 60 min. the primary rabbit polyclonal antibodies, SERPINE1 (Proteintech, United States), α-smooth muscle actin (α-SMA) (Proteintech, United States), β-catenin (Proteintech, United States), vimentin (HuaBio, China), ERK1/2 (HuaBio, China), phosphorylation extracellular signal-regulated kinase1/2 (p-ERK1/2) (HuaBio, China) and actin (ZSGB-BIO, China), and the mouse monoclonal antibody E-cad (HuaBio, China), were added to the membranes and incubated overnight at 4 °C.

Techniques: Expressing, Transfection, Migration, Marker

Journal: World Journal of Gastrointestinal Oncology

Article Title: MiRNA-145-5p inhibits gastric cancer progression via the serpin family E member 1- extracellular signal-regulated kinase-1/2 axis

doi: 10.4251/wjgo.v16.i5.2123

Figure Lengend Snippet: Effect of serpin family E member 1 overexpression on gastric cancer cell proliferation and the signal-regulated kinase-1/2 pathway in vitro and in vivo . A: Expression of signal-regulated kinase-1/2 (ERK1/2) and p-ERK1/2 in SGC7901 co-transfected cells; B: Image of xenograft tumors at day 18 post-inoculation; C: Xenograft tumor growth, volume, and mass at day-18 post-inoculation. n = 5 mice per group; D: Ki67 expression in xenograft tumor tissue; Scale bar = 20 μm; E: E-cadherin, vimentin, serpin family E member 1, ERK1/2, and p-ERK1/2 expression in xenograft tumor tissues. a P < 0.05; b P < 0.01; c P < 0.001. SERPINE1 : Serpin family E member 1; ERK1/2: Signal-regulated kinase-1/2.

Article Snippet: The proteins were then boiled at 100°C for 10 min with 100 μL of loading buffer (Beyotime Biotechnology, China) and separated by SDS-PAGE at 120 V. After electrophoresis, the proteins were transferred onto PVDF membranes and blocked using 5% BSA/TBST for 60 min. the primary rabbit polyclonal antibodies, SERPINE1 (Proteintech, United States), α-smooth muscle actin (α-SMA) (Proteintech, United States), β-catenin (Proteintech, United States), vimentin (HuaBio, China), ERK1/2 (HuaBio, China), phosphorylation extracellular signal-regulated kinase1/2 (p-ERK1/2) (HuaBio, China) and actin (ZSGB-BIO, China), and the mouse monoclonal antibody E-cad (HuaBio, China), were added to the membranes and incubated overnight at 4 °C.

Techniques: Over Expression, In Vitro, In Vivo, Expressing, Transfection