Journal: Biology

Article Title: Endothelial PAI-1 Drives Lead-Induced Cerebral Amyloid Angiopathy via Activation of C3 + Decorin + A1-like Astrocytes

doi: 10.3390/biology15040297

Figure Lengend Snippet: Pb exposure upregulates endothelial PAI-1 expression in vitro. ( A ) Representative Western blot showing PAI-1 protein expression in HUVECs treated with control (NaAc) or 1 µM PbAc. Uncropped Western blot images shown in supplement. ( B ) Quantification of PAI-1 protein levels in HUVEC lysates normalized to GAPDH ( n = 6). ( C ) PAI-1 concentrations in HUVEC-conditioned media (HUVEC-CM) with or without immunodepleting of PAI-1 or IgG control measured by ELISA. Data are demonstrated as mean ± SD; *: p < 0.05, analyzed by one-way ANOVA with Dunnett’s post hoc test. Note: Ctrl: control (sodium acetate), Pb: lead acetate, CM: conditioned media, IP: immunodepleting of PAI-1, PAI-1: plasminogen activator inhibitor-1.

Article Snippet: The concentration of PAI-1 in HUVEC- HUVEC-CM was determined using a human PAI-1 sandwich ELISA kit (E-EL-H2104, Elabscience, Houston, TX, USA) according to the manufacturer’s instructions.

Techniques: Expressing, In Vitro, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Biology

Article Title: Endothelial PAI-1 Drives Lead-Induced Cerebral Amyloid Angiopathy via Activation of C3 + Decorin + A1-like Astrocytes

doi: 10.3390/biology15040297

Figure Lengend Snippet: Depletion or inhibition of PAI-1 attenuates Pb-Induced Activation of C3 + Decorin + SVG Astrocytes in vitro. ( A ) Representative SVG astrocytes immunostained for GFAP (green), C3 (red), and decorin (magenta) after incubation with HUVEC-CM from Pb-treated HUVEC immunodepleted of PAI-1 or incubated with TIP. Arrowheads indicate positive C3 + decorin + A1-like astrocytes. ( B ) Quantification of triple-positive C3 + decorin + A1-like astrocytes. Data are demonstrated as mean ± SD, *: p < 0.05, **: p < 0.01, analyzed by one-way ANOVA with Dunnett’s post hoc test. Note: Pb: lead acetate, IP: immunodepleting of PAI-1, TIP: tiplaxtinin.

Article Snippet: The concentration of PAI-1 in HUVEC- HUVEC-CM was determined using a human PAI-1 sandwich ELISA kit (E-EL-H2104, Elabscience, Houston, TX, USA) according to the manufacturer’s instructions.

Techniques: Inhibition, Activation Assay, In Vitro, Incubation

Journal: Biology

Article Title: Endothelial PAI-1 Drives Lead-Induced Cerebral Amyloid Angiopathy via Activation of C3 + Decorin + A1-like Astrocytes

doi: 10.3390/biology15040297

Figure Lengend Snippet: Perivascular C3 + decorin + A1-like astrocytes in Pb-treated APP/PS1 mice with PAI-1 treatment. ( A ) Representative cortical sections from TIP-treated APP/PS1 mice with or without Pb exposure, triple-stained for GFAP (green), C3 (red), and decorin (magenta). Arrowheads indicate C3 + decorin + A1-like astrocytes surrounding cerebral vessels. ( B ) Quantification of C3 + decorin + A1-like astrocytes surrounding cerebral vessels ( n = 5 mice/group). Data are demonstrated as mean ± SD; *: p < 0.05, analyzed by one-way ANOVA with Dunnett’s post hoc test. Note: Pb: lead acetate, TIP: tiplaxtinin.

Article Snippet: The concentration of PAI-1 in HUVEC- HUVEC-CM was determined using a human PAI-1 sandwich ELISA kit (E-EL-H2104, Elabscience, Houston, TX, USA) according to the manufacturer’s instructions.

Techniques: Staining

Journal: Diabetes

Article Title: Plasminogen Activator Inhibitor-1 Is Involved in Streptozotocin-Induced Bone Loss in Female Mice

doi: 10.2337/db12-1552

Figure Lengend Snippet: Effects of PAI-1 deficiency on diabetic bone loss in both sexes of mice. Hematoxylin-eosin staining of tibia in control and streptozotocin-treated male and female PAI-1 WT and KO mice ( A ). BMD values in total, trabecular, and cortical bones ( B ); cortical thickness ( C ); and second moment of minimum and polar areas ( D ) of tibia in control and streptozotocin-treated male PAI-1 WT and KO mice. BMD values in total, trabecular, and cortical bones ( E ); cortical thickness ( F ); and second moment of minimum and polar areas ( G ) of tibia in control and streptozotocin-treated female PAI-1 WT and KO mice. For assessment of trabecular BMD, trabecular regions of interest extended from 96 um distal to the end of the proximal growth plate over 1.5 mm toward the diaphysis. For assessment of cortical BMD and thickness, cortical ROIs were defined as 2.0-mm segments of the mid-diaphysis tibia. For assessment of total BMD and bone strength index (second moment of minimum and polar areas: index of bending strength), ROIs were defined as 9,600-μm segment (100 slices) from distal end of proximal growth plate of tibia. Parameters used for the CT scans were as follows: tube voltage, 50 kVp; tube current, 500 μA; integration time, 3.6 ms; axial field of view, 48 mm; and voxel size of 48 × 96 μm with a slice thickness of 96 μm. Bone parameters were analyzed using the LaTheta software (version 3.40). Results are expressed as means ± SEM. * P < 0.05, ** P < 0.01 ( n = 5–7 in each group). Cont, control; STZ, streptozotocin.

Article Snippet: Plasma total PAI-1 was measured using a Murine Total PAI-1 ELISA kit (Molecular Innovations, Novi, MI).

Techniques: Staining, Software

Journal: Science signaling

Article Title: Extracellular signal–regulated kinase 5 promotes acute cellular and systemic inflammation

doi: 10.1126/scisignal.aaa3206

Figure Lengend Snippet: (A to P) Wild-type mice were treated intraperitoneally with XMD8-92 (50 mg/kg), XMD17-109 (50 mg/kg), BIX02189 (25 mg/kg), or vehicle [30% (2-hydroxypropyl)-β-cyclodextrin with or without 5% DMSO] 30 min before they were injected intravenously with Pam3Cys (2.5 mg/kg), LPS (10 mg/kg), or vehicle (0.9% saline). The plasma concentrations of IL-6, CCL2, and CCL3 and the activity of PAI-1 were quantified 2 hours after challenge. *P < 0.05 when comparing Pam3Cys- or LPS-treated mice in the presence or absence of inhibitor; #P < 0.05 when comparing untreated control mice with mice treated with Pam3Cys or LPS in the presence or absence of inhibitor. Data are means ± SD of four mice per group and are representative of two independent experiments.

Article Snippet: Active PAI-1 was detected with the PAI-1 functional assay ELISA kit (Molecular Innovations).

Techniques: Injection, Activity Assay

Journal: Science signaling

Article Title: Extracellular signal–regulated kinase 5 promotes acute cellular and systemic inflammation

doi: 10.1126/scisignal.aaa3206

Figure Lengend Snippet: (A to D) Wild-type mice were treated intraperitoneally with XMD8-92 (50 mg/kg) or vehicle [30% (2-hydroxypropyl)-β-cyclodextrin] 30 min before being injected intravenously with HKSA (2.5 × 1010 bacteria/kg) or vehicle (0.9% saline). The plasma concentrations of IL-6, CCL2, and CCL3 and the activity of PAI-1 were quantified 2 hours after the mice were challenged. *P < 0.05 when comparing HKSA-treated mice in the presence or absence of inhibitor; #P < 0.05 when comparing untreated control mice with mice treated with HKSA in the presence or absence of inhibitor. Data are means ± SD of four mice per group and are representative of two independent experiments.

Article Snippet: Active PAI-1 was detected with the PAI-1 functional assay ELISA kit (Molecular Innovations).

Techniques: Injection, Activity Assay