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. 2015 May 11;126(4):520–530. doi: 10.1182/blood-2014-09-601484

Figure 6.

Figure 6

Tmod3 regulates F-actin polymerization and organization in MKs. (A) Tmod3 is associated with F-actin bundles that assemble in MKs spread on collagen. Immunofluorescence staining of F-actin (red) and Tmod3 (green) in WT MKs spread on collagen for 30 minutes. F-actin polymerizes into bundles and foci on the ventral surface of the MKs. Boxed regions shown at high magnification (lower panels) reveal Tmod3 puncta associated with the F-actin bundles (yellow arrrowheads). Bars, 10 μm. (B-C) Immunofluorescence staining of F-actin (red) and NMIIA (green) in Tmod3+/+ and Tmod3−/− MKs spread on collagen. In WT MKs, F-actin assembles into parallel bundles, as well as intensely stained foci on the ventral surface, termed podosomes.17 NMIIA is associated with the F-actin bundles (but not the podosomes) in a striated pattern in WT MKs, whereas both F-actin and NMIIA form amorphous aggregates near the cell periphery in Tmod3−/− MKs. Boxed regions shown at higher magnification (lower sets of panels in [B-C]). Scale bars, 10 µm. Images are single optical sections from the ventral surface of MKs, acquired using a Zeiss LSM 780 confocal laser scanning fluorescence microscope with a ×63 oil-immersion objective (N.A. 1.4) at zoom 1 (top panels) and zoom 3 (boxed regions).