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. 2015 Mar 20;10(3):e0119739. doi: 10.1371/journal.pone.0119739

Fig 7. Restoration of collagen-induced platelet aggregation by enforced expression of PLCγ1 in PLCγ1/γ2 double-deficient platelets.

Fig 7

Lethally irradiated wild-type (WT) mice were reconstituted with WT bone marrow transfected with IRES-GFP retroviruses (WT/GFP) or with PLCγ1/γ2 double-deficient bone marrow transfected with IRES-GFP retroviruses (dKO/GFP), PLCγ1-IRES-GFP retroviruses (dKO/PLCγ1), or PLCγ2-IRES-GFP retroviruses (dKO/PLCγ2). Recipient mice were analyzed 8 weeks after reconstitution. (A) Overexpression of PLCγ1 or PLCγ2 in PLCγ1/γ2 double-deficient platelets. Washed non-sorted platelets from reconstituted recipients were subjected to direct Western blot analysis with antibodies specific for PLCγ1, PLCγ2 and Syk. (B) Washed non-sorted platelets from reconstituted recipients were stimulated under stirring conditions with collagen (6 μg/ml). Results were recorded on a Chrono-log Platelet Aggregometer. A representative aggregometry plot (adjusted for the timing of addition of collagen) is shown on the left, and quantitative analysis of the results of two independent experiments is shown on the right (*p < 0.005 for each genotype relative to dKO/GFP). Note that defective collagen-induced platelet aggregation was overcome by enforced expression of either PLCγ1 or PLCγ2 in PLCγ1/γ2 double-deficient platelets.