(A) Human platelets were incubated with PBS or candidalysin (CL) at 10 μM and with blocking reagents to the indicated platelet receptors as indicated after which secreted Dkk-1 was quantitated.
(B and C) Schematic diagrams and aggregate data depicting in vitro assays in which the dose-dependent binding of either plate-bound candidalysin or scrambled control peptide (SC) (B) or GP1bα (C) to the other reagent was determined colorimetrically (OD, optical density).
(D) Schematic diagram and aggregate data depicting in vitro binding assays with GP1ba blocking reagents (anti-GP1bα antibody; VWF A1A2A3 tridomain) in which the dose-dependent inhibition of binding of candidalysin is colorimetrically quantitated against plates coated with GP1bα.
(E) Pull-down assay of GP1ba from human platelet lysates using biotinylated candidalysin or SC as bait.
n ≥ 4, mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 using one-way ANOVA followed by Tukey’s test for multiple comparisons (A) or two-tailed Student’s t test (B–C). . Illustrative figures generated at biorender.com. Data are representative of two independent experiments.