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. 2016 Jun 21;291(32):16462–16476. doi: 10.1074/jbc.M116.730440

FIGURE 3.

FIGURE 3.

PKCϵ-mediated phosphorylation of PSD-95 at serine 295 is essential for its membrane accumulation. A, immunoblot representing p-PSD-95S295 and PSD-95 expression after incubation of the different combinations of recombinant PKCϵ, PSD-95, PKCϵ activators, and PKCϵ inhibitors (inh) mentioned above at 37 °C for 10 min in vitro. B, bryostatin 1 (Bry, 0.27 nm) and DCPLA-ME (100 nm) induced the phosphorylation of PSD-95 at serine 295 position. C, expression of p-PSD-95S295 and PSD-95 and β-actin in HEK-293 cells transfected with empty vector, wild type human PSD-95, and mutant PSD-95S295K. D, expression of PSD-95, p-PSD-95S295, PKCϵ, and β-actin in the soluble (S) and particulate (P) fraction of wild-PSD-95- and PSD-95S295K-transfected HEK-293 cells treated with bryostatin 1 and DCPLA-ME for 4 h in the presence or absence of EAVSLKPT (5 μm). S, soluble fractions; P, membrane fractions. Percentage of total protein in the membrane; PKCϵ (E), PSD-95 (F) and p-PSD-95S295 (G). Data are represented as the mean ± S.E. of three independent experiments (Student's t test. *, p < 0.05; **, p < 0.005; ***, p < 0.005).