Figure 2.
Quantitative analyses of Mks in situ.(Ai) High-volume 3D confocal image of E13.5 FL section showing Mks and Plts (anti-CD41, gray), and nuclei (DAPI, blue). Scale bar, 100 µm. (Aii) Surface objects rendered from raw data: Mks (gray); free Plts (yellow); nuclei pseudocolored according to DNA content (DAPI, color spectrum). Scale bar, 100 µm; n = 5 independent experiments. (Aiii) Enlargement of box: Mk with attached bud (arrow); free Plt (arrowhead). Scale bar, 20 µm. (Aiv) Optical section from raw data of budding Mk in iii; nuclear surface objects overlaid. Scale bar, 20 µm. (B) Quantitation of in vitro (i) and in vivo (ii) Plt-forming mechanisms (n = 4–5 independent experiments). P = 0.0002 (i) and P < 0.0001 (ii), unpaired two-tailed t test. (C) Diameters of circulating Plts (o = 236), free Plt in the FL (o = 406), and Mk-associated buds (o = 749; n = 3 independent experiments). P = 0.32, one-way ANOVA. (Di) Representative image of Plt-associated proteins in free Plt in E13.5 FL. Scale bar, 2 µm. (Dii) Representative image of Plt-associated proteins in Mk and associated buds in E13.5 FL. Scale bar, 2 µm. (Ei) Representative example (optical sections) of F-actin, MYH9, and β1-TUBULIN (β1-TUB) staining observed in free Plts in E13.5 FL. (Eii) Representative example of an optical section of a proPlt-forming Mk in E13.5 FL. (Eiii) Representative example of an optical section from a budding Mks in E13.5 FL showing the presence of a Plt-like pattern of MHY9 and β1-TUB staining in the membrane buds. Optical sections are taken from the boxed regions. Scale bars, 3 µm. n = six independent experiments. (F) In situ ploidy values for nonbudding Mks (i, o = 160), budding Mks (ii, o = 78), and proPlt-forming Mks (iii, o = 16); n = 5 independent experiments.

Quantitative analyses of Mks in situ. (Ai) High-volume 3D confocal image of E13.5 FL section showing Mks and Plts (anti-CD41, gray), and nuclei (DAPI, blue). Scale bar, 100 µm. (Aii) Surface objects rendered from raw data: Mks (gray); free Plts (yellow); nuclei pseudocolored according to DNA content (DAPI, color spectrum). Scale bar, 100 µm; n = 5 independent experiments. (Aiii) Enlargement of box: Mk with attached bud (arrow); free Plt (arrowhead). Scale bar, 20 µm. (Aiv) Optical section from raw data of budding Mk in iii; nuclear surface objects overlaid. Scale bar, 20 µm. (B) Quantitation of in vitro (i) and in vivo (ii) Plt-forming mechanisms (n = 4–5 independent experiments). P = 0.0002 (i) and P < 0.0001 (ii), unpaired two-tailed t test. (C) Diameters of circulating Plts (o = 236), free Plt in the FL (o = 406), and Mk-associated buds (o = 749; n = 3 independent experiments). P = 0.32, one-way ANOVA. (Di) Representative image of Plt-associated proteins in free Plt in E13.5 FL. Scale bar, 2 µm. (Dii) Representative image of Plt-associated proteins in Mk and associated buds in E13.5 FL. Scale bar, 2 µm. (Ei) Representative example (optical sections) of F-actin, MYH9, and β1-TUBULIN (β1-TUB) staining observed in free Plts in E13.5 FL. (Eii) Representative example of an optical section of a proPlt-forming Mk in E13.5 FL. (Eiii) Representative example of an optical section from a budding Mks in E13.5 FL showing the presence of a Plt-like pattern of MHY9 and β1-TUB staining in the membrane buds. Optical sections are taken from the boxed regions. Scale bars, 3 µm. n = six independent experiments. (F) In situ ploidy values for nonbudding Mks (i, o = 160), budding Mks (ii, o = 78), and proPlt-forming Mks (iii, o = 16); n = 5 independent experiments.

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