Figure 3.
Buds derive from the attached Mk.(Ai) Possible Pf4-Cre:Confetti cassette configurations after Cre-mediated recombination. (Aii) Flow cytometric analysis of E13.5 circulating Plts; note that mCFP is expressed only in configurations c and d Plts (histogram). (Aiii) Mk:bud match interpretation. (B) Optical section though configuration c (white arrow) and d (red arrow) budding Mks in E13.5 FL. Scale bar, 10 µm. (C) Mk:bud configuration scores (o = 732 buds, 184 Mks, n = 5 independent experiments). (Di) Representative example of a 3D z-projection of active CASPASE-3 (aCASP3) detection in fresh E13.5 FL (anti-aCASP3, blue; anti-CD41, gray). Cells undergoing apoptosis (aCASP3+) were rare. FL slices incubated with 4 µM Camptothecin (a potent inducer of apoptosis) were used as positive controls (n = 4 independent experiments). Scale bars, 50 µm. (Dii) Volume-normalized aCASP3 content of Plts and Mks compared with CD41− aCASP3+ objects (o = 2,100 Plts, 1,430 Mks, 139 CD41− aCASP3+ objects; n = 4 independent experiments). (Diii) Representative example of aCASP3 detection in situ, confirming the absence of CASPASE-3 activation in budding Mks. Optical section of budding Mk (boxed region); yellow arrow indicates a membrane-associated bud. Scale bars, 20 µm.

Buds derive from the attached Mk. (Ai) Possible Pf4-Cre:Confetti cassette configurations after Cre-mediated recombination. (Aii) Flow cytometric analysis of E13.5 circulating Plts; note that mCFP is expressed only in configurations c and d Plts (histogram). (Aiii) Mk:bud match interpretation. (B) Optical section though configuration c (white arrow) and d (red arrow) budding Mks in E13.5 FL. Scale bar, 10 µm. (C) Mk:bud configuration scores (o = 732 buds, 184 Mks, n = 5 independent experiments). (Di) Representative example of a 3D z-projection of active CASPASE-3 (aCASP3) detection in fresh E13.5 FL (anti-aCASP3, blue; anti-CD41, gray). Cells undergoing apoptosis (aCASP3+) were rare. FL slices incubated with 4 µM Camptothecin (a potent inducer of apoptosis) were used as positive controls (n = 4 independent experiments). Scale bars, 50 µm. (Dii) Volume-normalized aCASP3 content of Plts and Mks compared with CD41− aCASP3+ objects (o = 2,100 Plts, 1,430 Mks, 139 CD41− aCASP3+ objects; n = 4 independent experiments). (Diii) Representative example of aCASP3 detection in situ, confirming the absence of CASPASE-3 activation in budding Mks. Optical section of budding Mk (boxed region); yellow arrow indicates a membrane-associated bud. Scale bars, 20 µm.

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