Critical role of PLCγ2 in MK differentiation, migration, and spreading. (A) DNA ploidy was analyzed using flow cytometry by staining purified BM-derived mature MKs from WT and PLCγ2-deficient mice with propidium iodide and FITC-CD41 antibody. Representative profiles and quantification of the percentage of cells with differing levels of ploidy and the modal ploidy from 3 independent experiments are shown. **P < .01; ***P < .005. (B) Purified BM-derived mature MKs from wild-type (WT) and PLCγ2 knockout (PLCγ2 KO) were exposed to a SDF1α gradient over 3 hours within the Dunn chamber and the net translocation distance of each cell was measured *** P < .005. (C) CXCR4 surface expression of purified BM-derived mature MKs from WT and PLCγ2-deficient mice (PLCγ2 KO) was analyzed by flow cytometry. Gray line indicates relevant control antibody; black line, FITC-conjugated anti-CXCR4 antibody. Representative profiles and quantification of the percentage of cells expressing CXCR4 from 3 independent experiments are shown. (D) Purified BM-derived mature MKs from WT and PLCγ2-deficient mice (PLCγ2 KO) were plated on fibronectin-coated surface for 3 hours. Adherent MKs were fixed and permeabilized and actin fibers stained with rhodamine-phalloidin. Representative images (scale bar = 20μm) and surface area quantification from 4 independent experiments are shown. **P < .01. (E) Purified BM-derived mature MKs from WT and PLCγ2-deficient mice (PLCγ2 KO) were plated on fibronectin (FN) or maintained in suspension in a BSA-coated dish (BSA) for 3 hours. MKs were lysed and WCLs were analyzed by Western blot with MLC-P and MLC antibodies. Blots are representative of 3 independent experiments.