Fig. 6: MICAL1 promotes stable platelet adhesion to VWF by increasing resistance to shear-induced morphological changes. | Nature Communications

Fig. 6: MICAL1 promotes stable platelet adhesion to VWF by increasing resistance to shear-induced morphological changes.

From: F-actin disassembly by the oxidoreductase MICAL1 promotes mechano-dependent VWF-GPIbα interaction in platelets

Fig. 6: MICAL1 promotes stable platelet adhesion to VWF by increasing resistance to shear-induced morphological changes.

a, b Whole blood from Mical1+/+ or Mical1−/− mice was perfused over a recombinant mouse VWF-coated surface at 1500 s⁻¹. a Representative time-lapse images showing individual platelet tether formation. Red arrows indicate tethers. Scale bars, 2 µm. See Supplementary Movies 4 and 5. b Tether elongation was quantified as [(length at 3 min/length at 0 s) ×100]; mean ± SD, N = 30 tethers from 3 independent experiments (Welch’s t-test, t = 4.105, df = 31.52). c Tether length was tracked over time; N = 30 tethers from 3 experiments; mean ± 95% CI (two-way ANOVA with Šídák’s test, F(9, 504) = 4.375; p = 0.003 at 80 s; p < 0.001 from 100 to 160 s). d–f Unstained Mical1+/+ or Mical1−/− blood was perfused on VWF. Post-perfusion, coverslips were fixed and stained with phalloidin and anti-integrin β3. d Platelet surface area measured in 3 experiments (273 platelets/experiment); mean ± SD (Student’s t test, t = 3.938, df = 4). e F-actin quantification in tethers at 1500 s⁻¹ using phalloidin (Mical1+/+ = 39, Mical1−/− = 37 tethers; Welch’s t test, t = 3.261, df = 61.62). Left: representative images with actin-positive tethers (red arrows). Scale bars, 1 µm. f Tether length measured from at least 254 platelets/experiment (N = 3); mean ± SD (two-way ANOVA with Fisher’s LSD test, F(1,8) = 22.09). Left: representative β3-labeled tethers at 1500 and 9000 s⁻¹. Scale bars, 5 µm. g Scanning electron microscopy (SEM) images of platelets from 9000 s⁻¹ flow assays. Circularity index quantified in 2 experiments (n = 2 mice/condition pooled); Mical1+/+ = 65, Mical1−/− = 63 platelets (Student’s t test, t = 4.495, df = 126). Scale bars: 4 µm (overview), 1–2 µm (zoom). h Frequency of “ghost” platelets per field assessed by SEM in 2 experiments (Mical1+/+ = 743, Mical1−/− = 609 platelets; Student’s t test, t = 4.495, df = 126). Representative SEM images with zooms shown. Scale bars: 4 µm and 2 µm.Violin plots display medians (central line) and interquartile ranges (25th–75th percentiles). ns: not significant. Source data are provided as a Source Data file.

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