G-protein Inhibitor reduces Pls-mediated phosphorylation of Akt and ERK in the neuronal cells.

<p>(A) Pls-mediated phosphorylation of ERK and Akt was cancelled by treatments with the G-protein inhibitor. The neuronal cells (N2A) were cultured in low serum (2% FBS) containing medium for 20 hr and then treated with the G-protein inhibitor GDPβS at 200 μM concentration for 4 hours followed by Pls (500 ng/ml) treatments for 20 minutes. Cell extracts were then subjected to western blotting assays. (B) The quantification data of panel A showed that Pls treatment failed to increase phosphorylation of ERK and Akt in the GDPβS pretreated cells (One way ANOVA followed by Bonferoni’s test, n = 5, *, <i>P</i><0.05 and **, <i>P</i><0.01. n.s. stands for not significant). Image-J software was used to quantify the signals. (C) Real-time PCR data shows the orphan GPCRs expression in the primary neurons, astrocytes, microglia and also in the cell lines of N2A (neuronal derived cells), A1 (astrocytes) and MG6 (microglia). The data represent mean ± S.E.M (standard error of mean) (n = 3).