Fig 2: ACTB-AST patients display minor facial anomalies and thrombocytopenia with enlarged platelets. a Craniofacial appearance of patient 3 (left, P3, p.Ala331Val_fs*27) at 5 years of age, patient 4 (mid, P4, p.Ala331Val_fs*27) at 31 years of age and patient 5 (right, P5, p.Ser338_Ile341del) at 4 year 10 months. Flared eyebrows (P3 and P4), straight eyebrows (P4 and P5), telecanthus (all), epicanthal folds (P3 and P5), upslanting palpebral fissures (P3 and P4), a broad nasal tip (P3 and P5), a bulbous nose (P4), a thin upper vermillion border (all) and prominent chin (P4) are observed in these patients; b, c CD61-labeled platelets purified from a healthy control (C), P3, P4 and P5 were analyzed by immunofluorescence microscopy; b Representative images show that platelets in patient samples vary from normal to large in size. Scale bars represent 5 µm; c Particle analysis shows a significant shift in the size distribution and average diameter of patient platelets compared with a healthy control. Individual data points are plotted with the median and IQR. The number of platelets analyzed from 1 experiment is given in brackets below each condition. Significance was determined with the Kruskal–Wallis and Dunn’s multiple comparisons tests, where ****p < 0.0001

Fig 3: Optical images of platelets and platelet-derived microvesicles (PMVs). Platelets obtained from PRP (platelet-rich plasma) were placed on siliconized (a) and collagen-coated (b, c) glass slides. Phalloidin staining revealed specific actin cytoskeleton organization. Thrombin activation produced more phalloidin-stained spots (c). Arrows indicate actin streaks from activated platelets, which correspond to actin-rich PMVs. ImageStreamX Mark II cytometry showed that most of the PMVs were gated between 0.13 and 0.22 μm in sidescatter (SSC) and bright-field channels (d). A minority (∼30 %) of the PMVs were positive for annexin V (e); most of the PMVs were double-positive for the antigens CD9 (exosome-specific) and CD61 (platelet-specific) (f). Selected images of microvesicles are shown in g. AF Alexa Fluor, PE phycoerythrin

Fig 4: Heterogeneity of CD61 expression in Circulating Tumor Cells (CTCs) from prostate cancer patients. Confocal microscopy images of CTCs positive for CK (FITC.green) and nucleus (DAPI. blue). CD61 platelet marker absent and present in CTCs (AF-647. red). Abbreviations: CK: cytokeratin, AF: Alexa Fluor.

Fig 5: Phenotypic characterization and evaluation of antigen uptake by immature dendritic cells (DCs). Monocytes cultured in presence or absence of activated platelets were differentiated into immature DCs using IL4 and GM-CSF (50 ng/ml each) for 5 d. (1A and 1B) Scanning electron microscopy and transmission electron microscopy images of platelet-DC complexes. The platelet is marked by a black star symbol. (1C–1H) DCs that were derived from monocytes cocultured with platelets and expressed CD61, a platelet marker, are termed “Plt.” DCs that were derived from monocytes cocultured with platelets, did not express CD61, but were still exposed to platelet-derived biomolecules are called “Plt Releasates.” DCs derived from noncomplexed monocytes (not treated; NT), and CD40L treated monocytes (CD40L) were used as controls. Cells from each group were stained with respective monoclonal antibodies and analyzed using Accuri C6 flow cytometer and FlowJo software v10. The data is shown as raw data or fold change as compared to NT (HIV uninfected individuals, n = 6–9, 4 males and 5 females). Results were analyzed by repeated measures 1-way ANOVA followed by Tukey's multiple comparisons test using GraphPad prism v7. * indicates P < 0.05, **indicates P < 0.01, and ***indicates P < 0.00. Percentages and mean fluorescence intensity (MFI) for expression of (1C) CD163, (1D) CD206, (1E) DC-SIGN, (1F) CD16, and (1G) CCR5 on DCs using Accuri C6 flow cytometer. (1H) MFI for phagocytosed IgG-Fluoresbrite microspheres by immature DCs