FIGURE 5.

Lysophosphatidic acid receptor5 (LPAR5) activates PI3K kinase through p110β catalytic isoform. A, Lysophosphatidic acid(LPA) increased PIP3 generation. CGTH‐W3 cells transfected with pEGFP‐C1‐Grp1‐PH were incubated in serum‐free media for 24 h. After addition of LPA (10 μM) for 5 min, the fluorescent images of the cell were captured with confocal microscopy. B, LPAR5 associates with p110β catalytic isoform in thyroid cancer cells. In immunoprecipitation assay, the pulling antibody was LPAR5. Control immunoprecipitation was performed using rabbit IgG. Protein samples from whole‐cell lysates were used to point at the position of the protein examined. IP, immunoprecipitated; IB, immunoblot. C, D, E, The expression of phosphorylated Akt and p70S6K induced by LPA is lower in the cells transfected with p110β‐siRNA. After transfection with the respective siRNA for 24 h, CGTH‐W3 and TPC‐1 cells were maintained in FBS‐free medium and then stimulated with LPA (10 μM) for 10 min. Phosphorylated Akt and p70S6K were analyzed by immunoblotting; GAPDH was employed as loading control