Fig. 5: Zyxin deficiency affects microtubule and actin cytoskeleton organization.

A Left panels, representative confocal images of resting WT, Zyx^−/− and Gp1ba^−/− platelets stained for β-tubulin (green). Scale bar: 3 μm. Right panel, the quantification of MFI of β-tubulin on the left was analyzed by ImageJ software; n = 10 visual fields from five mice per genotype. Data are expressed as means ± SD. B Confocal images of WT and Zyx^−/− platelets were allowed to spread on fibrinogen in the presence of thrombin and stained for F-actin (green). Scale bar: 3 μm. Images are representative of five independent experiments. C, D MKs cultured from mouse FL HPCs were allowed to spread on type I collagen. Representative confocal images of spread WT and Zyx^−/− MKs stained for F-actin (green) and nuclei (blue); scale bar: 10 μm (C). The percentage of MKs with the organized F-actin network along collagen fibers (as indicated by arrows in C) (D). No less than 60 MKs per genotype from seven independent experiments were analyzed. E, F MKs cultured from mouse FL HPCs were allowed to spread on fibrinogen. Representative confocal images of spread WT and Zyx^−/− MKs stained for F-actin (green) and nuclei (blue); scale bar: 10 μm (E). The percentage of MKs with lamellipodia (as indicated by arrows in E) (F). No less than 60 MKs per genotype from eight independent experiments were analyzed (C–F). The original magnification of all the images is ×630. Means are indicated by horizontal lines in (D) and (F). ***P < 0.001, compared with WT mice by one-way ANOVA followed by Dunnett’s post hoc test in (A) and by unpaired Student’s t-test in (D) and (F).