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. 2014 Aug 6;124(14):2271–2279. doi: 10.1182/blood-2014-05-575878

Figure 6.

Figure 6

12-LOX modulates early signaling components of the FcγRIIa pathway in human platelets. Washed human platelets were treated with ML355 (20 μM) or vehicle control prior to stimulation by crosslinking (IV.3 + GAM). (A) A time course of PLCγ2 phosphorylation at site Y759 was assessed by western blot analysis. All samples were normalized to total PLCγ2 and fold changes were quantified relative to the unstimulated condition. The bar graph comprised of n = 7 individuals. (B) Following crosslinking (IV.3 + GAM), calcium mobilization was measured by flow cytometry. Representative curves were quantitated in fold change of free calcium relative to the unstimulated condition over 4 minutes. Bar graphs represent the ratio of the fold change in calcium mobilization (n = 5). (C) N = 7 stimulated human platelets with or without ML355 were analyzed for PKC activity following CD32 crosslinking, and PMA rescue comprised of n = 3. A PKC substrate was blotted as a surrogate for PKC activation and pleckstrin phosphorylation. Data represents mean ± SEM. *P < .05; **P < .01.