Effects of EPA, DHA and Gö6976 on ANXA2 on the cell surface and in culture supernatant, and on Ser-phosphorylation of ANXA2.

<p>Cells were pretreated with Hank's-EGTA and examined after culturing for 24 h. (A) EPA (10 µM) reduced, and DHA (10 µM) increased the amount of 36 kDa and also 33 kDa in the EGTA extract (upper lane). In the culture supernatant (lower lane), these PUFAs marginally reduced the amount of 36 kDa. (B) The cells were incubated EPA and Gö6976 (50 nM). Gö6976 increased the surface-bound 36 kDa. It also suppressed the effect of EPA. (C) Effects of EPA, DHA and OLA on Ser-phosphorylation of ANXA2. The same protein amounts of whole extract of cells treated with indicated fatty acids (10 µM) were incubated with anti-phosphoserine mAb and the same portions of immunoprecipitates were examined for ANXA2. EPA increased the amount of the Ser-phosphorylated ANXA2. In contrast, DHA slightly reduced the amount.