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. 2020 Mar 11;295(16):5307–5320. doi: 10.1074/jbc.RA119.012296

Figure 6.

Figure 6.

Calcium-dependent generation of 12-HETE-lysophospholipids in murine platelets isolated from WT and iPLA2γ−/− (KO) mice stimulated with calcium ionophore A23187. Washed platelets were incubated in the absence or presence of the calcium ionophore A23187 (1.0 μm) at 37 °C for 15 min prior to extraction of lipids using a modified Bligh and Dyer (56) procedure and analysis by HPLC-MS/MS. For inhibition experiments, ML355 (10 μm) was preincubated for 15 min prior to addition of A23187. A and B, content of 2-AA-LPC (A) and 2-AA-LPE (B) in the absence (DMSO) (Control) or presence (A23187) of calcium ionophore A23187 or ML355 and A23187 (ML355-A23187) in platelets from WT or KO mice. C and D, content of 12-HETE-LPC (C) and 12-HETE-LPE (D) in the absence (DMSO) (Control) or presence (A23187) of calcium ionophore A23187 or ML355 and A23187 (ML355-A23187) in platelets from WT or KO mice. Data are expressed as mean ± S.E. of six independent replicates. Statistical analyses were performed using an unpaired two-tailed Student's t test. Statistically significant p values are as indicated above the bracketed data sets (**, p < 0.01; ***, p < 0.001; ****, p < 0.0001).