Journal: Scientific Reports

Article Title: Tissue Plasminogen Activator Neurotoxicity is Neutralized by Recombinant ADAMTS 13

doi: 10.1038/srep25971

Figure Lengend Snippet: ( A ) Representative immunoblots and quantitative analysis of the calpain- (145/150 kDa) and caspase-specific fodrin cleaved products (120 kDa) in sham-operated mice, and mice treated with vehicle, tPA, tPA in combination with rADAMTS 13, and rADAMTS 13 24 hours after MCA occlusion. The gels have been run under the same experimental conditions. ( B ) Six fields selected for immunohistochemical quantification of LC3, TUNEL, and FJB-labeled cells in the peri-infarct area in Nissl-stained coronal sections in all the groups. ( C ) Representative photomicrographs of TUNEL staining and quantification of TUNEL-positive cells in the ischemic brain of mice treated with vehicle, tPA, rADAMTS 13 in combination with tPA, and rADAMTS 13 24 hours after MCA occlusion. ( D ) Representative photomicrographs of Fluoro-Jade B (FJB) staining and quantification of FJB-positive cells in the ischemic brain of mice treated with vehicle, tPA, rADAMTS 13 in combination with tPA, and rADAMTS 13 24 hours after MCA occlusion. ( E ) Representative immunoblots of LC3 and quantification of the LC3-II/LC3-I ratio in sham-operated mice, and mice treated with vehicle, tPA, tPA in combination with rADAMTS 13, and rADAMTS 13 24 hours after MCA occlusion. The gels have been run under the same experimental conditions. ( F ) Representative photomicrographs of LC3 staining and quantification of LC3-positive cells in the ischemic brain of mice treated with vehicle, tPA, rADAMTS 13 in combination with tPA, and rADAMTS 13 24 hours after MCA occlusion. Bar = 30 μm. Values are mean ± SD (n = 5 per group). *P < 0.05.

Article Snippet: The primary antibodies were rabbit anti-LC3 (Light Chain 3, 4108), rabbit anti-pERK1/2 (phosphorylation of extracellular signal-regulated kinase1/2, 9101), rabbit anti-pJNK (c-Jun N-terminal kinases, 9251), rabbit anti-p-p38 (9211), rabbit anti-NR1 (N-methyl-D-aspartate receptor, 4204), rabbit anti-NR2A (4205), rabbit anti-NR2B (4207), rabbit anti-β-actin (4967) (all from Cell Signaling Technology, Beverly, MA, USA), mouse anti-α-fodrin (AA6, Enzo Life Sciences, Inc. Farmingdale, NY, USA), rabbit anti-tPA (ASHTPA-GF-HT, Molecular Innovations Inc., Novi, MI, USA).

Techniques: Western Blot, Immunohistochemical staining, TUNEL Assay, Labeling, Staining

Journal: Circulation

Article Title: Releasing the Brakes on the Fibrinolytic System in Pulmonary Emboli: Unique Effects of Plasminogen Activation and α2-Antiplasmin Inactivation

doi: 10.1161/CIRCULATIONAHA.116.024421

Figure Lengend Snippet: Lung tissue containing FITC-fibrin labeled (green) pulmonary emboli was immunostained (red) to detect the expression of (A) tPA, (B) uPA and (C) plasminogen (Pg) (D) The total immune-stained area (arbitrary units; a.u) for each protein was measured in a 20× (100 μm) image of an embolized vs non embolized (Control; dashed yellow outline) pulmonary artery in the lungs. n=3, mean ± SEM. **p<0.01; ns, non-significant.

Article Snippet: The primary antibodies include rabbit anti-mouse against tPA (ASMTPA-GF; Molecular Innovations), uPA (urokinase plasminogen activator) (ASMUPA-GF-HT; Molecular Innovations) and PAI-1 (IASMPAI-GF; Innovative Research), rabbit anti-mouse plasminogen (Abcam), goat anti-mouse α2-antiplasmin (AF1239; R&D Systems), and rat anti-mouse Ly6G (clone1A8, #127602; Biolegend).

Techniques: Labeling, Expressing, Staining