Figure 3.
HSPCs polarize upon interaction with osteoblasts.(A) Representative confocal images of an HSPC cultured on an osteoblast in a microwell. Upper panel: Tilted 3D view. Lower panel: Lateral view. Actin appears in red, microtubules in green, DNA in blue, and centrosome in white. The HSPC is polarized, with its centrosome proximal to the point of contact with the osteoblast, highlighted with an arrow, as in all images. Scale bars = 5 µm. (B) Golgi organization in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: Golgi appears in red, actin in green, centrosome in white, and DNA in blue. Lower panel: Inverted image of the Golgi. Scale bar = 5 µm. (C) Microtubule organization in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: Microtubules appear in red, actin in green, centrosome in white, and DNA in blue. Lower panel: Inverted image of the microtubules. Scale bar = 5 µm. (D) Actin cytoskeleton organization in a polarized HSPC. Selected Z stacks of representative cells are presented. In all images, microtubules are in green and DNA in blue. First column: Actin appears in red in the upper panel, and the lower panel contains an inverted image of Actin. Second column: Arp2/3 appears in red in the upper panel, and the lower panel contains an inverted image of Arp2/3. Third column: Ezrin appears in red in the upper panel, and the lower panel contains an inverted image of Ezrin. Fourth column: phospho-myosin light chain appears in red in the upper panel and in black in the lower panel (inverted image). Scale bars = 5 µm. (E) Localization of CD133 in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: CD133 appears in red, actin in green, microtubules in white, and DNA in blue. Lower panel: Inverted image of CD133. Scale bar = 5 µm. (F) Localization of CD44 in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: CD44 appears in red, actin in green, centrosome in white, and DNA in blue. Lower panel: Inverted image of CD44. Scale bar = 5 µm. (G) Graphical summary of the localization of the factors described in a polarized HSPC.

HSPCs polarize upon interaction with osteoblasts. (A) Representative confocal images of an HSPC cultured on an osteoblast in a microwell. Upper panel: Tilted 3D view. Lower panel: Lateral view. Actin appears in red, microtubules in green, DNA in blue, and centrosome in white. The HSPC is polarized, with its centrosome proximal to the point of contact with the osteoblast, highlighted with an arrow, as in all images. Scale bars = 5 µm. (B) Golgi organization in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: Golgi appears in red, actin in green, centrosome in white, and DNA in blue. Lower panel: Inverted image of the Golgi. Scale bar = 5 µm. (C) Microtubule organization in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: Microtubules appear in red, actin in green, centrosome in white, and DNA in blue. Lower panel: Inverted image of the microtubules. Scale bar = 5 µm. (D) Actin cytoskeleton organization in a polarized HSPC. Selected Z stacks of representative cells are presented. In all images, microtubules are in green and DNA in blue. First column: Actin appears in red in the upper panel, and the lower panel contains an inverted image of Actin. Second column: Arp2/3 appears in red in the upper panel, and the lower panel contains an inverted image of Arp2/3. Third column: Ezrin appears in red in the upper panel, and the lower panel contains an inverted image of Ezrin. Fourth column: phospho-myosin light chain appears in red in the upper panel and in black in the lower panel (inverted image). Scale bars = 5 µm. (E) Localization of CD133 in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: CD133 appears in red, actin in green, microtubules in white, and DNA in blue. Lower panel: Inverted image of CD133. Scale bar = 5 µm. (F) Localization of CD44 in a polarized HSPC. Selected Z stacks of a representative cell. Upper panel: CD44 appears in red, actin in green, centrosome in white, and DNA in blue. Lower panel: Inverted image of CD44. Scale bar = 5 µm. (G) Graphical summary of the localization of the factors described in a polarized HSPC.

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