Journal: Science Advances

Article Title: PAI-1 is a critical regulator of FGF23 homeostasis

doi: 10.1126/sciadv.1603259

Figure Lengend Snippet: Plasma levels of cFGF23 (black filled bars) and iFGF23 (gray filled bars) were measured in WT, untreated, and TM5441-treated Pai-1stab mice ( A ) and in WT, kl/kl , TM5441-treated kl/kl , and kl/klpai-1 −/− mice ( B ). As seen in (A), changes in cFGF23 levels due to PAI-1 overexpression and TM5441 treatment were both significant in Pai-1stab mice. Similarly, TM5441 treatment and PAI-1 deficiency significantly reduced cFGF23 and iFGF23 levels in kl/kl mice. # P = 0.0003, $ P = 0.0001 versus kl/kl cFGF23; * P = 0.03, ** P = 0.001 versus kl/kl iFGF23. Analysis of the measurements by one-way analysis of variance (ANOVA) yielded significant differences ( P < 0.0001) for both Pai-1stab and kl/kl mice groups.

Article Snippet: Purified recombinant human two-chain tPA (catalog no. HTPA-TC), uPA (catalog no. UPA-LMW), and active PAI-1 (catalog no. CPAI) were obtained from Molecular Innovations.

Techniques: Over Expression

Journal: Science Advances

Article Title: PAI-1 is a critical regulator of FGF23 homeostasis

doi: 10.1126/sciadv.1603259

Figure Lengend Snippet: ( A ) AKI induces plasma PAI-1 levels 10-fold higher in FA-treated WT mice ( n = 13) compared to vehicle-treated WT mice ( n = 5). ( B ) Mouse iFGF23 (miFGF23) levels in WT ( n = 7), WT with AKI ( n = 21), TM5441-treated WT with AKI ( n = 15), PAI-1 −/− with AKI ( n = 9), and pai-1 −/− untreated ( n = 6). * P = 0.007, # P = 0.005. ( C ) Effect of PAI-1 inhibition on clearance of rhFGF23 in WT AKI mice ( n = 7) and TM5441 (10 mg/kg per day)–treated WT AKI mice ( n = 7). The rhFGF23 was injected into the tail vein, and human cFGF23 (hcFGF23) levels were measured in plasma samples collected at time points 1, 3, 5, 15, 30, 60, and 120 min. ( D ) The cFGF23 levels at 1 min were considered as 100% to plot percent remaining values of rhFGF23 over the time course of the experiment.

Article Snippet: Purified recombinant human two-chain tPA (catalog no. HTPA-TC), uPA (catalog no. UPA-LMW), and active PAI-1 (catalog no. CPAI) were obtained from Molecular Innovations.

Techniques: Inhibition, Injection

Journal: Science Advances

Article Title: PAI-1 is a critical regulator of FGF23 homeostasis

doi: 10.1126/sciadv.1603259

Figure Lengend Snippet: TM5441 treatment or PAI-1 deficiency did not affect the increased serum phosphate levels ( # P < 0.05 versus WT) ( A ) and elevated plasma levels of PTH (* P < 0.01 versus WT) ( B ) in kl/kl and FA-treated WT and pai-1 −/− mice. mPTH, mouse PTH.

Article Snippet: Purified recombinant human two-chain tPA (catalog no. HTPA-TC), uPA (catalog no. UPA-LMW), and active PAI-1 (catalog no. CPAI) were obtained from Molecular Innovations.

Techniques:

Journal: Science Advances

Article Title: PAI-1 is a critical regulator of FGF23 homeostasis

doi: 10.1126/sciadv.1603259

Figure Lengend Snippet: ( A ) Silver-stained SDS-PAGE gel of reactions containing purified recombinant human tPA, PAI-1, and FGF23 with and without the PAI-1 antagonist TM5441. ( B ) Model depicting that inhibition of PAI-1 leads to restoration of PA- and furin-mediated FGF23 proteolysis.

Article Snippet: Purified recombinant human two-chain tPA (catalog no. HTPA-TC), uPA (catalog no. UPA-LMW), and active PAI-1 (catalog no. CPAI) were obtained from Molecular Innovations.

Techniques: Staining, SDS Page, Purification, Recombinant, Inhibition

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