Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: SOX2-OT induced by PAI-1 promotes triple-negative breast cancer cells metastasis by sponging miR-942-5p and activating PI3K/Akt signaling

doi: 10.1007/s00018-021-04120-1

Figure Lengend Snippet: SOX2-OT activates PI3K/Akt signaling pathway via miR-942-5p. a Putative miR-942-5p binding site and mutant sequences in the 3’UTR of PIK3CA. b Luciferase activity of PIK3CA WT and PIK3CA MUT upon transfection of miR-942-5p mimics in 293 T cells. ***P < 0.001 by t test. c PIK3CA mRNA level of MDA-MB-231 and Hs578t transfected with miR-942-5p mimic or inhibitor. ***P < 0.001 by t test. d Relative protein levels of MDA-MB-231 and Hs578t transfected with miR-942-5p mimic or inhibitor. e, f Wound-healing assay (e) and Transwell invasion assay (f) of MDA-MB-231 and Hs578t cells that were treated with co-transfection of miR-942-5p mimic and PIK3CA WT or MUT vector. Photographs were obtained immediately (0 h) and at 48 h after wounding. *P < 0.05, **P < 0.01, ***P < 0.001 by t test. g, h PIK3CA mRNA level of MDA-MB-231 and Hs578t transfected with SOX2-OT WT, SOX2-OT MUT or shSOX2-OT, and miR-942-5p mimic or inhibitor. ***P < 0.001 by t test. i Relative protein levels of MDA-MB-231 and Hs578t transfected with SOX2-OT WT, SOX2-OT MUT or shSOX2-OT, and miR-942-5p mimic or inhibitor. Results represented the average of three independent experiments, the data represent the mean ± SD

Article Snippet: Human recombinant PAI-1 (Topscience #T4254, Shanghai, China) were dissolved in the culture medium, PAI-1 inhibitor PAI-039 (Topscience #T2030, Shanghai, China), PI3K inhibitor LY294002 (Topscience #T2008, Shanghai, China) were dissolved in dimethylsulfoxide (DMSO, Sigma) and stored at − 80 °C as a 1 μg/ml stock solution.

Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Transfection, Wound Healing Assay, Transwell Invasion Assay, Cotransfection, Plasmid Preparation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: SOX2-OT induced by PAI-1 promotes triple-negative breast cancer cells metastasis by sponging miR-942-5p and activating PI3K/Akt signaling

doi: 10.1007/s00018-021-04120-1

Figure Lengend Snippet: Schematic model of PAI-1/SOX2-OT/miR-942-5p/PI3K/Akt signaling in the TNBC metastasis. PAI-1 promotes SOX2-OT expression. In patients with a low PAI-1 expression, SOX2-OT blinds to miR-942-5p and the expression level of SOX2-OT is also low. In patients with a high PAI-1 expression, SOX2-OT acts as a molecular sponge for miR-942-5p, leading to activating PI3K/Akt signaling pathway and promoting TNBC invasion and metastasis

Article Snippet: Human recombinant PAI-1 (Topscience #T4254, Shanghai, China) were dissolved in the culture medium, PAI-1 inhibitor PAI-039 (Topscience #T2030, Shanghai, China), PI3K inhibitor LY294002 (Topscience #T2008, Shanghai, China) were dissolved in dimethylsulfoxide (DMSO, Sigma) and stored at − 80 °C as a 1 μg/ml stock solution.

Techniques: Expressing

Journal: In Vivo

Article Title: Inhibition of Plasminogen Activator Inhibitor-1 (PAI-1) by Tiplaxtinin Attenuates the Aggressive Phenotype of Vulvar Squamous Cell Carcinoma Cells In Vitro

doi: 10.21873/invivo.14345

Figure Lengend Snippet: Pronounced PAI-1 signals are detectable in both A431 and Cal-39 VSCC cells. Detection of PAI-1 protein expression in three different passages of A431 and Cal-39 VSCC cells using western blot. PAI-1: Plasminogen Activator Inhibitor-1; VSCC: vulvar squamous cell carcinoma.

Article Snippet: The cells were treated with PAI-1 inhibitor tiplaxtinin (PAI-039, TPX) (MedChemExpress, Monmouth Junction, NJ, USA) dissolved in 2% v/v ethanol (Th.

Techniques: Expressing, Western Blot

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Plasminogen Activator Inhibitor-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells

doi: 10.1161/ATVBAHA.124.320938

Figure Lengend Snippet: Human coronary artery SMCs (passage number 5) were treated with vehicle control or PAI-039 (10μM) for 24 hours, after which cortical stiffness (elastic modulus) was assessed by atomic force microscopy. Results of 9–10 independent experiments for each treatment are shown with mean±SEM for each group. P value was determined with the Mann-Whitney U test.

Article Snippet: PAI-039, a specific inhibitor of PAI-1, 18 was from Pfizer, Cambridge, MA, USA.

Techniques: Inhibition, Control, Microscopy, MANN-WHITNEY

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Plasminogen Activator Inhibitor-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells

doi: 10.1161/ATVBAHA.124.320938

Figure Lengend Snippet: (A) Human SMCs (passage number 5–7) were treated with PAI-039 (2.5–10μM) or vehicle control for 18 hours after which z-stack images were acquired by confocal microscopy and F/G-actin ratio was quantified. Results of 3 independent experiments for each PAI-039 concentration are shown with mean±SEM of each group. P values shown were determined by one-way repeated measures ANOVA with Geisser-Greenhouse correction and Tukey’s post hoc testing. (B) Representative images of human SMCs. Arrows indicate stress fibers. (C, D) Murine SMCs (passage number 5) were treated with PAI-039 (5μM) or vehicle control for 18 hours, after which F/G actin ratio was determined. Results of 5 independent experiments with wild-type SMCs (C) and 4 independent experiments with PAI-1-knockout SMCs (D) are shown with mean±SEM for each experimental group. P values shown were determined with the Mann-Whitney U test. (E) Representative images of murine SMCs. Scale bars represent 25 μm; F-actin is red; G-actin is green; nuclei are blue.

Article Snippet: PAI-039, a specific inhibitor of PAI-1, 18 was from Pfizer, Cambridge, MA, USA.

Techniques: Inhibition, Control, Confocal Microscopy, Concentration Assay, Knock-Out, MANN-WHITNEY

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Plasminogen Activator Inhibitor-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells

doi: 10.1161/ATVBAHA.124.320938

Figure Lengend Snippet: Smooth muscle cells (SMCs, passage number 5–7) were treated with PAI-039 for 18 hours in serum-poor conditions (0.2% fetal bovine serum), then cell lysates were prepared and the ratio of phospho (P)-cofilin to total cofilin was quantified by SDS-PAGE and western blotting. (A) Results of 6 independent experiments with human SMCs treated with PAI-039 (10μM) or vehicle control. (B) Results of 7 independent experiments with wild-type murine SMCs treated with PAI-039 (5μM) or vehicle control. (C) Results of 7 independent experiments with PAI-1-knockout murine SMCs treated with PAI-039 (5μM) or vehicle control. In (A-C), numerical data include mean±SEM for each experimental group, a representative western blot is shown to the right of the graph, and P values shown were determined with the Mann-Whitney U test.

Article Snippet: PAI-039, a specific inhibitor of PAI-1, 18 was from Pfizer, Cambridge, MA, USA.

Techniques: Activation Assay, SDS Page, Western Blot, Control, Knock-Out, MANN-WHITNEY

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Plasminogen Activator Inhibitor-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells

doi: 10.1161/ATVBAHA.124.320938

Figure Lengend Snippet: Human smooth muscle cells (passage number 7) were treated with PAI-039 (10μM) or vehicle control for 0, 0.5, 1, or 3 hours, after which cell lysates were prepared and ratios of phosphorylated (P)-to-total protein were quantified by SDS-PAGE and western blotting for (A) cofilin, (B) LIM domain kinase 1 (LIMK) and (C) slingshot homolog 1 (SSH). All data points shown are mean±SEM of 3 independent experiments. P values shown were determined by two-way repeated measures ANOVA with Geisser-Greenhouse correction and Sidak multiple comparison post hoc testing. Representative western blots accompany each graph.

Article Snippet: PAI-039, a specific inhibitor of PAI-1, 18 was from Pfizer, Cambridge, MA, USA.

Techniques: Activation Assay, Control, SDS Page, Western Blot, Comparison

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Plasminogen Activator Inhibitor-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells

doi: 10.1161/ATVBAHA.124.320938

Figure Lengend Snippet: (A, B) Human coronary artery smooth muscle cells (SMCs, passage number 7) were serum-starved for 1 hour, then exposed to 4 experimental conditions, generated by adding dorsomorphin (4μM) or vehicle control for 30 minutes, then adding PAI-039 (10μM) or vehicle control for 60 minutes. Cell lysates were prepared, and ratios of phosphorylated (P)-to-total (A) AMPK and (B) cofilin were quantified by SDS-PAGE and western blotting with results normalized to the negative control group not treated with dorsomorphin or PAI-039. Results of 5 independent experiments, including mean±SEM of each group, are shown. (C) Representative western blots of human SMC lysates. (D) Murine SMCs isolated from wild-type or PAI-1-knockout (PAI-1-KO) mice (passage number 5) were serum starved for 1 hour, then treated for 1 hour with PAI-039 (5μM) or vehicle control. Cell lysates were prepared, and ratios of P-to-total AMPK were quantified by SDS-PAGE and western blotting with results normalized to wild-type SMCs not treated with PAI-039. Results of 4 independent experiments, including mean±SEM of each group, are shown. (E) Representative western blots of murine SMC lysates. For all graphs, P values shown were determined with the Kruskal-Wallis test with uncorrected Dunn’s test post hoc testing.

Article Snippet: PAI-039, a specific inhibitor of PAI-1, 18 was from Pfizer, Cambridge, MA, USA.

Techniques: Generated, Control, SDS Page, Western Blot, Negative Control, Isolation, Knock-Out

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Plasminogen Activator Inhibitor-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells

doi: 10.1161/ATVBAHA.124.320938

Figure Lengend Snippet: Age and sex-matched wild-type mice were fed a western diet containing or lacking PAI-039. (A) After 5 weeks, pulse wave velocity (PWV) in the abdominal aorta was significantly reduced in animals receiving PAI-039 (n=7 mice) vs . controls (n=7 mice). (B-H) Histologic studies. After 8.5 weeks, mice were euthanized and cross-sections from each abdominal aorta were stained for collagen and elastin content. There was no significance between groups in (B) tunica media thickness, (C) elastin content of the tunica media, or (E) collagen content of the full-thickness aortic wall. Representative images of (D) elastin (black) and (F) collagen (brick red) staining are shown. Scale bars represent 50 μm. (G) F-actin content of the tunica media was significantly lower in PAI-039-treated mice than controls. (H) Representative confocal fluorescence microscopy images of F-actin staining (red); nuclei are blue; L, lumen; scale bars represent 10 μm. All graphs include mean±SEM of each group. P values shown were determined with the Mann-Whitney U test (A and C) or the unpaired student t-test (B, E, and G).

Article Snippet: PAI-039, a specific inhibitor of PAI-1, 18 was from Pfizer, Cambridge, MA, USA.

Techniques: Inhibition, Western Blot, Staining, Fluorescence, Microscopy, MANN-WHITNEY