Journal:

Article Title: Selectin-mucin interactions as a probable molecular explanation for the association of Trousseau syndrome with mucinous adenocarcinomas

doi: 10.1172/JCI200318882

Figure Lengend Snippet: Time-course of platelet-rich microthrombi in wild-type mouse lungs following a single injection of mucin. (a) Typical representations of in vivo platelet-rich microthrombi in lung sections observed using fluorescently labeled Ab against the platelet marker, CD41. (b) Quantitative histologic determination, quantitating average number of CD41-positive pixels per 200× view field (mean value of 20 random fields). Each bar represents the mean ± SEM. *P < 0.001 compared with PBS-injected mice, n = 7; **P < 0.05, n = 5.

Article Snippet: Acetone-fixed cryosections (5 μm) were blocked in 10% rabbit serum, incubated with sheep anti-human fibrin(ogen) (BIODESIGN International Inc., Saco, Maine, USA) (PBS, 1% BSA, 0.1% NaN 3 , 4°C, overnight), FITC-conjugated rabbit anti-sheep IgG heavy & light (H&L) chains, purified rat anti-mouse CD41 (BD PharMingen), AffiniPure Cy 3 -conjugated donkey anti-rat IgG (H&L) (Jackson ImmunoResearch Laboratories Inc., West Grove, Pennsylvania, USA), and mounted with Vectashield mounting medium containing DAPI (Vector Laboratories Inc., Burlingame, California, USA).

Techniques: Injection, In Vivo, Labeling, Marker

Journal:

Article Title: Selectin-mucin interactions as a probable molecular explanation for the association of Trousseau syndrome with mucinous adenocarcinomas

doi: 10.1172/JCI200318882

Figure Lengend Snippet: Selectin deficiency attenuates platelet-rich microthrombus formation following mucin injection. (a) Typical representations of platelet-rich microthrombi in lung sections observed as in Figure ​Figure2,2, at 5 minutes after injection. (b) Quantitative determination of CD41 in lung sections as in Figure ​Figure2.2. Values shown represent the mean ± SEM. *P < 0.001 compared with mucin-injected wild-type mice, n = 7; **P < 0.05, n = 7. White bars, PBS-injected mice; black bars, mucin-injected mice.

Article Snippet: Acetone-fixed cryosections (5 μm) were blocked in 10% rabbit serum, incubated with sheep anti-human fibrin(ogen) (BIODESIGN International Inc., Saco, Maine, USA) (PBS, 1% BSA, 0.1% NaN 3 , 4°C, overnight), FITC-conjugated rabbit anti-sheep IgG heavy & light (H&L) chains, purified rat anti-mouse CD41 (BD PharMingen), AffiniPure Cy 3 -conjugated donkey anti-rat IgG (H&L) (Jackson ImmunoResearch Laboratories Inc., West Grove, Pennsylvania, USA), and mounted with Vectashield mounting medium containing DAPI (Vector Laboratories Inc., Burlingame, California, USA).

Techniques: Injection

Journal:

Article Title: Selectin-mucin interactions as a probable molecular explanation for the association of Trousseau syndrome with mucinous adenocarcinomas

doi: 10.1172/JCI200318882

Figure Lengend Snippet: Relative importance of fluid-phase coagulation versus selectins in mucin-induced microthrombus formation. (a and b) Quantitative determination of CD41 in lung sections as in Figure ​Figure2,2, at 5 minutes after injection. (a) Ability of heparin or hirudin to block the effects of mucin in wild-type mice. *P < 0.05 compared with mucin alone, n = 5; **P < 0.001 compared with PBS-injected mice, n = 5. (b) Effects of coinjecting subthreshold levels of thrombin with mucin. (c) Three-color immunofluorescence microscopic analysis of platelet-rich microthrombi in wild-type mice injected with mucin (with or without hirudin) to demonstrate fibrin, an indicator of thrombin activity. Bright green indicates deposited fibrin; pale green indicates autofluorescence of luminal surface of longitudinally sectioned venule; red indicates platelets; and blue indicates nuclei. Examples of occluding microthrombi and diffuse platelet aggregates are shown.

Article Snippet: Acetone-fixed cryosections (5 μm) were blocked in 10% rabbit serum, incubated with sheep anti-human fibrin(ogen) (BIODESIGN International Inc., Saco, Maine, USA) (PBS, 1% BSA, 0.1% NaN 3 , 4°C, overnight), FITC-conjugated rabbit anti-sheep IgG heavy & light (H&L) chains, purified rat anti-mouse CD41 (BD PharMingen), AffiniPure Cy 3 -conjugated donkey anti-rat IgG (H&L) (Jackson ImmunoResearch Laboratories Inc., West Grove, Pennsylvania, USA), and mounted with Vectashield mounting medium containing DAPI (Vector Laboratories Inc., Burlingame, California, USA).

Techniques: Coagulation, Injection, Blocking Assay, Immunofluorescence, Activity Assay