Neuronal Orphan G-Protein Coupled Receptor Proteins Mediate Plasmalogens-Induced Activation of ERK and Akt Signaling
Fig 5
The Pls specific cellular signaling mediated by the GPCRs.
(A) Western blotting assays showed the reduction of p-ERK and p-Akt expression in the N2A cells where the Pls-synthesizing enzyme GNPAT was knocked down by sh-GNPAT lentivirus. The data represent three independent experiments. (B) The quantification data of the panel A showed the significant reduction of phosphorylated ERK and Akt proteins in the GNPAT knockdown cells. The data represent mean ± S.E.M (Student’s t-test, n = 3, **, P < 0.01). (C) Serum deprived N2A cells were treated with Pls (500 ng/ml) and PtdEtn (phosphatidylethanolamine, 500 ng/ml) for 20 minutes. Cells extracts were then subjected to western blotting assays to detect phosphorylation status of ERK and Akt proteins. The data represent three independent experiments. (D) Quantification data of panel C showed that PtdEtn treatments did not significantly increase the phosphorylated ERK and Akt as compared with the Pls treatments group. The data represent mean ± S.E.M (Bonferroni’s test; n = 3, *, P < 0.05). (E) N2A cells were infected by sh-GNPAT and the control sh-Luc lentivirus particles for 24 hours flowed by the overexpression of the GPCRs expressing plasmids for 48 hours. Cell extracts were then subjected to western blotting assays. The data represents three independent experiments (n = 3). (F) Quantification data of the panel E showed that the GPCRs-mediated increase in phosphorylation of ERK in the control lentivirus group (sh-Luc) was abolished in the sh-GNPAT groups. The data represent mean ± S.E.M (Dunnett`s test; n = 3, *, P < 0.05).