PI(3,4,5)P3 analog stimulates detyrosination of microtubules.

<p><b>[A]</b> NIH/3T3 cells were siRNA depleted for GAPDH (control) or PTEN and serum depleted overnight. Cell lysates were analyzed by western blotting against PTEN, phospho-FAK (Y397), total FAK, phospho-PDK1, phospho-Akt (Ser473), total Akt, phospho-GSK3β, total GSK3β and β-Actin. Representative images of 3 independent experiments are shown. <b>[B, C]</b> NIH/3T3 cells were siRNA depleted against GAPDH or PTEN, treated with FAK inhibitors (PF-562271[1 μM], PF-573228 [10 μM]) as indicated and seeded on fibronectin-coated coverslips for 2 h. <b>[B]</b> Representative images of cells immunostained against, detyrosinated tubulin (red), total tubulin (green) and DNA (blue). <b>[C]</b> Percentage of detyrosinated microtubules positive cells. Error bar = StdDev, N = 4 (total of >150 cells each). <b>[D, E]</b> NIH/3T3 cells were serum depleted for 2 days and treated with LPA [10 μM] (positive control) or PIP4-AM as indicated. <b>[D]</b> Representative image of cells immunostained against detyrosinated tubulin (deTyr, red), total tubulin (green) and DNA (blue). <b>[E]</b> Percentage of deTyr positive cells. Error bar = StdDev, N = 3 (total of >150 cells each). Scale bars, 10 μm, ** p<0.01, *** p<0.001.