Journal: Journal of Thrombosis and Haemostasis

Article Title: Cleavage by MMP‐13 renders VWF unable to bind to collagen but increases its platelet reactivity

doi: 10.1111/jth.14729

Figure Lengend Snippet: Inhibition of platelet adhesion, rolling, and thrombus formation on intact and cleaved von Willebrand factor (VWF) using anti‐αIIbβ3 and anti‐GpIbα compounds. A, Binding of washed platelets to intact and cleaved VWF in xCELLigence solid phase binding assays. Collagen type I and bovine serum albumin (BSA) were used as control substrates. * P < .05, ** P < .01 (one‐way analysis of variance [ANOVA] and post‐hoc Tukey honestly significant difference [HSD] test). Where indicated, washed platelets were pre‐incubated with 1 or 10 µmol/L anti‐αIIbβ3 and/or 10 or 50 µg/mL anti‐GpIbα compounds. B, Whole blood platelet rolling at a shear rate of 1000 seconds on intact (green symbols) and cleaved (red symbols) VWF, respectively. Where indicated, washed platelets were pre‐incubated with 10 µmol/L anti‐αIIbβ3 and/or 50 µg/mL anti‐GpIbα compounds. C, Results are shown for (i) surface coverage, (ii) mean height, and (iii) ZV50. Results are mean values of four separate donors ± SE

Article Snippet: The slides were blocked with 1% BSA for 1 hour prior to the addition of 1:1000 dilution anti‐human GpIbα (AF4067; R&D Systems, Abingdon) or anti‐P‐selectin (ab6632; abcam) overnight at 4°C.

Techniques: Inhibition, Binding Assay, Control, Incubation, Shear

Journal: Journal of Thrombosis and Haemostasis

Article Title: Cleavage by MMP‐13 renders VWF unable to bind to collagen but increases its platelet reactivity

doi: 10.1111/jth.14729

Figure Lengend Snippet: GpIbα and P‐selectin expression on platelets following whole blood flow experiments. Whole blood was drawn through a flow chamber for 5 minutes over intact or cleaved von Willebrand factor (VWF) using a syringe pump to generate a wall shear rate of 1000 seconds -1 , corresponding to arteriolar conditions. Adherent platelets were then fixed in formalin prior to GpIbα and P‐selectin detection using FITC Alexa‐647 conjugated secondary antibodies respectively. A, Representative images of protein expression. B, Overall fluorescence and fold change of GpIbα and P‐selectin expression were calculated using Mean Gray value in ImageJ1.35. C, Fluorescent surface area of GpIbα and P‐selectin expression were determined following thresholding of images in ImageJ1.35. Data are the mean ± SE of nine separate donors. ** P < .005 (two‐tailed paired t ‐test)

Article Snippet: The slides were blocked with 1% BSA for 1 hour prior to the addition of 1:1000 dilution anti‐human GpIbα (AF4067; R&D Systems, Abingdon) or anti‐P‐selectin (ab6632; abcam) overnight at 4°C.

Techniques: Expressing, Shear, Fluorescence, Two Tailed Test

Journal: Cell

Article Title: Elevated Calprotectin and Abnormal Myeloid Cell Subsets Discriminate Severe from Mild COVID-19

doi: 10.1016/j.cell.2020.08.002

Figure Lengend Snippet:

Article Snippet: CD42 , Fluidigm , Pro# 3144020B.

Techniques: Recombinant, Reverse Transcription, SYBR Green Assay, Staining, Functional Assay, Software