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Blood, Vol. 111, Issue 7, 3407-3414, April 1, 2008
A nonsynonymous SNP in the ITGB3 gene disrupts the conserved membrane-proximal cytoplasmic salt bridge in the  IIbβ3 integrin and cosegregates dominantly with abnormal proplatelet formation and macrothrombocytopenia
Blood Ghevaert et al.
111: 3407
Supplemental materials for: Ghevaert et al
Files in this Data Supplement:
- Figure 1 (JPG, 97.6 KB)
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(A) Phase contrast microphotographs of CHO transfectants seeded for 2 hours at 37°C on fibrinogen (FIB) or serum proteins. Arrowheads point to the bulbous tips of the extending processes. Scale bar, 40 µm. (B) Immunofluorescence staining of CHO  IIb 3-D723 and -H723 cells seeded on a fibrinogen-coated surface. Tubulin was visualized using the -tubulin antibody B-512 Staining of the IIb3-H723 CHO cells showed a typical pattern of longitudinally oriented bundled microtubules emerging from the centrosome and extending to the end of the protrusions (B), similar to that seen in proplatelets in adherent MKs (C panel b). In wild-type cells, they radiated randomly from the centrosome towards the plasma membrane (B). Actin was visualised with phalloidin-rhodamine and showed continuous membrane staining along the protrusions, similar to the actin membrane lining observed in adherent MKs (B and C panel c). Arrowhead indicates the tip of the protrusion where the typical dense organised actin skeleton present in the distal MK proplatelet tip (C panel d) also seen in circulating platelets (C panel e) was not visible. Scale bar, 10 µm. (C) Light microscopy (a) and immunofluorescence (b-e) of CD34+ derived MKs (a-d) and platelets (e) plated on fibrinogen for 3 hours at 37C and stained for tubulin and actin.). Panel (d) is an enlargement of the proplatelet distal tip (white box, panel c). Scale bars 40 µm (a); 5 µm (b, c); 2,5 µm (d, e).
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