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