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: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 1. Upregulation of ALDH1A3 in oxGBM cells is associated with the increased expression and release of pro-angiogenic factors PAI-1 and IL-8. GBM cell lines were transduced with ALDH1A3 for overexpression (oxGBM) or with empty vector (evGBM). (A) Confirmation of up-regulation of ALDH1A3 mRNA level in oxGBM cells by RT2-PCR. (B) Confirmation of up-regulation of ALDH1A3 protein expression by Western blot. wt, wild-type cells. The uncropped blots are shown in Figure S7; (C) Angiogenesis array. The blots showed duplicated dots for 55 angiogenesis-related proteins in the media of oxU373 or evU373. 10 of 55 proteins were upregulated more than 2-fold in ox group compared to ev group (indicated by rectangle). They are: (1) Ang-1, (2) artemin, (3) TF, (4) ET-1, (5) GM-CSF, (6) IL-8, (7) PDGF-AA, (8) PAI-1, (9) PEDF, and (10) uPA. (D) Semi-quantification of the dots representing PAI-1 and IL-8. (E) Immunofluorescence staining of GBM cells. U373 (left panel) and LN229 (right panel). Co-localization of ALDH1A3 with PAI-1 and IL-8 was observed in oxGBM cells, whereas no immunoreactivity of PAI-1 and IL-8 was detected in evGBM cells. (F) mRNA expression of PAI-1 and IL-8 in transduced U373 cells and the effect of inhibitors. Tiplaxtinin (Tip, 30 µM) and reparixin (Rep, 1 µM) are the specific inhibitors of PAI-1 and IL-8 receptors CXCR1/2, respectively. (G) Detection of PAI-1 and potential signaling proteins by Western blot. IOD: optical density. The uncropped blots are shown in Figure S8. **, p < 0.01; ***, p < 0.001, compared with ev. ##, p < 0.01; ###, p < 0.001, compared with ox.

Article Snippet: To determine the effects of GBM-derived PAI-1 and IL-8 on EC behavior, the following inhibitors were used: Tiplaxtinin (cat# HY-15253; MedChemExpress, Monmouth Junction, NJ, USA), a small-molecule inhibitor of PAI-1, and reparixin (cat# HY-15251; Med-ChemExpress), an allosteric inhibitor of the IL-8 receptors CXCR1 and CXCR2.

Techniques: Expressing, Transduction, Over Expression, Plasmid Preparation, Western Blot, Staining

Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 2. Overexpression of ALDH1A3 in GBM cells activated endothelial angiogenesis in indirect co-culture with endothelial cells (ECs), which was reversed by treatment with respective inhibitors of PAI-1 or IL-8 receptors CXCR1/2. Indirect co-culture was performed by culture of HBMECs in a conditioned medium (CM) containing the media derived from evGBMs or oxGBMs and ECGM in a ratio of 1:1. PAI-1 inhibitor tiplaxtinin (Tip, 30 µM) and CXCR1/2 inhibitor reparixin (Rep, 1 µM) or vehicle DMSO (0.1%) was added to CM, followed by EC behavior study. All data were reproduced in three independent experiments. (A) Proliferation assay in HBMEC and HUVEC. Indirect co-culture of HBMECs and HUVECs with CM derived from oxU373 and oxLN229 stimulated EC proliferation, which was completely reversed by the treatment of tiplaxtinin, not by reparixin. (B) Scratch assay in HBMEC. Left panel: images were acquired 24 h after scratching. Scale bar: 200 µm. Right panel: quantitative analysis. Culture of HBMECs with CM derived from oxU373 or oxLN229 (oxCM) significantly promoted HBMEC migration, which was reversed by the treatment of tiplaxtinin and reparixin, respectively. (C) Transwell invasion assay in HBMEC. Left panel: Representative images of invaded cells were acquired after 24 h of incubation. Scale bar: 100 µm. Right panel: quantitative analysis. Culture of HBMECs with oxCM accelerated HBMEC invasion. This effect was significantly inhibited by the treatment of reparixin but not by tiplaxtinin. (D) Tube formation assay in HBMEC. Left panel: representative images of tube formation. Scale bar: 200 µm. Right panel: quantitative analysis of branching points per field. Tube formation in HBMECs was stimulated by incubation with oxCM, which was completely diminished by both inhibitors. (E) Sprouting assay in HBMEC. Left panel: representative images of sprouting in HBMECs after 24 h of co-culture. Scale bar: 100 µm. A pronounced increase in sprouting was observed in HBMECs cultured in oxCM. Tiplaxtinin and reparixin suppressed the sprouting effect resulting from oxCM. *, p < 0.05; **, p < 0.01 and ***, p < 0.001, compared with evCM. #, p < 0.05; ##, p < 0.01 and ###, p < 0.001, compared with oxCM.

Article Snippet: To determine the effects of GBM-derived PAI-1 and IL-8 on EC behavior, the following inhibitors were used: Tiplaxtinin (cat# HY-15253; MedChemExpress, Monmouth Junction, NJ, USA), a small-molecule inhibitor of PAI-1, and reparixin (cat# HY-15251; Med-ChemExpress), an allosteric inhibitor of the IL-8 receptors CXCR1 and CXCR2.

Techniques: Over Expression, Co-Culture Assay, Derivative Assay, Proliferation Assay, Wound Healing Assay, Migration, Transwell Invasion Assay, Incubation, Tube Formation Assay, Cell Culture

Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 3. Overexpression of ALDH1A3 in GBM cells stimulated EC proliferation, migration, and sprouting in direct co-culture with endothelial cells (ECs), which was inhibited by treatment with the specific inhibitors of PAI-1 or IL-8 receptors CXCR1/2. Direct co-culture was performed by co- culture of CellTrace™CFSE pre-labeled HBMECs with transduced U373 and LN229 in a ratio of 1:1 in ECGM. To inhibit PAI-1, tiplaxtinin (Tip, 30 µM) was pretreated with GBM cells 6 h in advance. For IL-8 receptor inhibition, reparixin (Rep, 1 µM) was pretreated with ECs 30 min in advance and added to ECGM in the co-culture, as well. All data were reproduced in three independent experiments. (A) Representative images of directly co-cultured CellTrace™CFSE-labeled HBMEC (green) with transduced U373 and LN229. Scale bar: 100 µm. (B) HBMEC proliferation. Direct co-culture of oxGBMs with HBMECs promoted the proliferation of HBMEC, which was completely reversed by the treatment of tiplaxtinin and reparixin. The fluorescence intensity of CFSE-labeled HBMECs, reflecting cell proliferation, was detected 48 h after direct co-culture at 485 nm of excitation wavelength and 535 nm of emission wavelength. (C) HBMEC migration. The images were acquired after 12 h of direct

Article Snippet: To determine the effects of GBM-derived PAI-1 and IL-8 on EC behavior, the following inhibitors were used: Tiplaxtinin (cat# HY-15253; MedChemExpress, Monmouth Junction, NJ, USA), a small-molecule inhibitor of PAI-1, and reparixin (cat# HY-15251; Med-ChemExpress), an allosteric inhibitor of the IL-8 receptors CXCR1 and CXCR2.

Techniques: Over Expression, Migration, Co-Culture Assay, Labeling, Inhibition, Cell Culture

Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 5. Immunohistochemistry and immunofluorescence staining on GBM sections. (A,B) Im- munohistochemistry staining of ALDH1A3. The immunoreactivity of ALDH1A3 was detected in vessels (arrows in (A)) and peripheral cells of glomeruloid (arrowheads in (B)) as well as in tumor cells (asterisks in (A)). Scale bar: 50 µm in (A) and 20 µm in (B). (C–E) Double-immunofluorescence staining of ALDH1A3 (red) with PAI-1 (green) (C,D) or with IL-8 (green) (E,F). The co-localization of ALDH1A3 with PAI-1 or with IL-8 immunoreactivity was detected in microvessels (arrows), glomeruloid (arrowheads), and tumor cells (asterisks) in GBM sections.

Article Snippet: To determine the effects of GBM-derived PAI-1 and IL-8 on EC behavior, the following inhibitors were used: Tiplaxtinin (cat# HY-15253; MedChemExpress, Monmouth Junction, NJ, USA), a small-molecule inhibitor of PAI-1, and reparixin (cat# HY-15251; Med-ChemExpress), an allosteric inhibitor of the IL-8 receptors CXCR1 and CXCR2.

Techniques: Immunohistochemistry, Staining

Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 6. Schematic illustration of the pro-angiogenic function of ALDH1A3 via paracrine PAI-1 and

Article Snippet: To determine the effects of GBM-derived PAI-1 and IL-8 on EC behavior, the following inhibitors were used: Tiplaxtinin (cat# HY-15253; MedChemExpress, Monmouth Junction, NJ, USA), a small-molecule inhibitor of PAI-1, and reparixin (cat# HY-15251; Med-ChemExpress), an allosteric inhibitor of the IL-8 receptors CXCR1 and CXCR2.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Effect of AZ3976 on clot lysis in human plasma with added PAI-1. Before addition of plasma, 1.7 nm PAI-1 in buffer was preincubated briefly at 37 °C with 0 to 0.22 mm AZ3976. The reaction was started by dilution with plasma containing tPA to final concentration of 50% plasma, 1.2 nm tPA and 0.65 nm PAI-1. Fibrin formation was monitored from the change in A405 in a microplate reader at 2-min intervals at 37 °C for up to 10 h. A, example of plasma clot lysis experiment: A405 is plotted versus time. Curves are shown for PAI-1 with 82.5 (b), 41.25 (c), 20.6 (d), 10.3 (e), 5.16 (f), and 0 μm (g) AZ3976 and for tPA only (a), without added PAI-1. B, the IC50 determination. The clot lysis time was defined as the time to halve the amplitude on the negative slope minus the time to halve the amplitude on the positive slope where the maximal effect (100%) was the time without added PAI-1 and minimal effect (0%) the time without added compound. The line depicts the best fit using Grafit IC50 fit. The mean of four experiments is plotted ± S.E.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Lysis, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Titration of glycosylated active and latent PAI-1 by isothermal calorimetry with AZ3976. Titration of 45 μm active PAI-1 with specific activity ∼75%, or 50 μm glycosylated latent PAI-1 in 50 mm sodium phosphate (pH 7.4), 100 mm NaCl, and 2% DMSO was achieved by injecting 20 × 0.5 μl and 20 × 1 μl aliquots, respectively, of 1 mm AZ3976 at 35 °C with 90 s waiting time between subsequent injections. The line depicts the best fit according to a single site binding model used to extract the thermodynamic properties of the interactions, which are shown in Table 2.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Titration, Activity Assay, Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: The structure of latent PAI-1 in complex with AZ3976. A, comparison of the latent PAI-1·AZ3976 holostructure with the latent PAI-1 apoform (PDB code 1DVN) (18). PAI-1 is colored by sequence from blue (N-terminal) to red (C-terminal). The inhibitor is shown in stick representation. The structure of the latent PAI-1 apostructure is shown as a gray ribbon representation. B, comparison of the AZ3976 binding site in ligand bound and free latent PAI-1. C, AZ3976 binding site in latent PAI-1. The Fo − Fc map calculated using the refined AZ3979-bound structure of latent PAI-1 is contoured at 3σ. D–F show the surface of the ligand binding site in three different forms of PAI-1. D, latent ligand complexed (this work); E, latent (PDB code 1DVN) (18); and F, active (PDB code 1DVM) (18). The magenta-colored surface describes the solvent accessible area in the region of AZ3979 binding. The nomenclature for the secondary structure is adopted from Stein and Chotia (50).

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Sequencing, Binding Assay, Ligand Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Interaction of PAI-1 with immobilized VN and effect of AZ3976. A, sensorgram overlay of the association and dissociation in real time after injection of 10 nm PAI-1 with 10 μm AZ3976 (a), incubated for increasing time intervals (arrow from 0 to 215 min) at 20 °C >900 RU immobilized VN, at change to buffer in the presence of 10 μm AZ3976 (b), and at injection of 20 μl of 25 nm tPA in the presence of 10 μm AZ3976 (c), followed by running buffer (d). The sensorgram of buffer only (dashed line) and reference, 10 nm PAI-1 in buffer without preincubation (marked ref) are included, whereas the sensorgrams of PAI-1 incubated in buffer are not shown for clarity. The ΔRU between the channel with immobilized VN, flow channel 2, and the channel without VN, flow channel 1, is shown. B, data are from the experiment shown in A. Initial binding rate to VN, measured after 10 s, as a function of the incubation time for 10 nm PAI-1 in buffer and for 10 nm PAI-1 with AZ3976 are shown. The lines are fitted to the data assuming single exponential decay, k = 0.0007 (buffer) and 0.01 min−1 (AZ3976), respectively.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Injection, Incubation, Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Interaction of PAI-1 with captured tPA and effect of incubation time with AZ3976. A, initial rate, measured as ΔRU, the change in RU 10 s after injection over tPA, as a function of the incubation time (min) of 10 nm PAI-1 with AZ3976 at different concentrations at 20 °C. Lines are drawn by fitting a single exponential decay to the data. B, rate constants from the data in A (circles) for single exponential decay of PAI-1, or from the data in Fig. 5 (triangles) from binding to VN, versus the concentration of AZ3976. The line was drawn by a linear fit with a slope of 14 m−1·s−1.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Incubation, Injection, Binding Assay, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Decay of PAI-1 activity and concomitant generation of latent PAI-1. A, sensorgram overlay of injection onto the monoclonal antibody H4B3 (specific for non-glycosylated latent PAI-1) captured on immobilized RamFc at 10 nm non-glycosylated active PAI-1 in buffer (dotted line) or with 20 μm AZ3976 for 1 h (solid line), or buffer alone (dashed line) at 20 °C (a). b, flow change to running buffer with 20 μm AZ3976. The ΔRU between the channel with H4B3 and the channel with the reference antibody is shown. B, values in % of ΔRU for 10 nm PAI-1, measured from the sensorgram 70 s after injection, which represent active PAI-1 bound to captured tPA and latent PAI-1 bound to H4B3, without AZ3976 and with 20 μm AZ3976 incubated with PAI-1 for 1 h at 20 °C.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Activity Assay, Injection, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: SPR sensorgrams for the interaction of AZ3976 with immobilized glycosylated latent and active PAI-1. Concentrations ranged from 20 μm to 27.4 nm following a 3-fold dilution scheme. The SPR experiments were run in PBSTD buffer at 25 °C, using a flow rate of 30 μl/min. The normalized responses were obtained by dividing the observed response levels with the predicted maximum response levels for a 1:1 binding interaction. The kinetic analysis was performed by fitting the data to a 1:1 binding model (fitted model traces as red dotted line). A, sensorgrams of AZ3976 binding to latent PAI. The normalized responses indicate a ligand binding activity of ∼90%. B, sensorgrams of AZ3976 binding to active PAI. The normalized responses indicate a ligand binding activity of ∼30%.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Binding Assay, Ligand Binding Assay, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Overview of inhibitor binding to PAI-1. A, the structure of active PAI-1 in complex with the somatomedin B domain of VN (PDB code 1OC0). PAI-1 is shown in a gray ribbon representation, except for the RCL, which is colored red (gap due to missing residues in structure). Somatomedin B is shown as a ribbon diagram in magenta. Additional residues implicated in VN binding are shown in stick representation colored in magenta and yellow. Residues implicated in binding of the small molecule PAI-1 inhibitor PAI-039 are shown in cyan and yellow, i.e. residues shown in yellow are implicated in both PAI-039 and VN binding. Glycosylation sites are shown in blue stick representation, and additional residues mentioned in the discussion are shown in green. B, the structure described in this work. The overall structure is colored the same way as in A, highlighting the different topology of the central β-sheet in the two structures. AZ3976 is shown in an orange stick representation and residues within 5 Å of the inhibitor are shown as cyan sticks.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: Binding Assay

Journal: The Journal of Biological Chemistry

Article Title: Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation *

doi: 10.1074/jbc.M112.371732

Figure Lengend Snippet: Mechanistic scheme of PAI-1 in presence of latency accelerating inhibitor. A represents active PAI-1, pL represents prelatent PAI-1, L represents latent PAI-1, and x indicates inhibitor.

Article Snippet: Active non-glycosylated human PAI-1 expressed in Escherichia coli was purchased from Molecular Innovations, Inc. (Novi, MI) and was shown to have a specific activity of 78%.

Techniques: