FIGURE 5.

3-MA inhibits class I and class III PI3K in different temporal patterns. A, the effect of 3-MA and wortmannin (Wort) on production of PI3P. WT MEFs were treated in EBSS or with 3-MA (5 mm), wortmannin (50 nm) in full medium as indicated. The levels of PI3P were measured using the PI3P mass strip kit as described under “Experimental Procedures.” B, the effect of 3-MA and wortmannin on production of PI(3,4,5)P₃. WT MEFs were treated as in A, and the levels of PI(3,4,5)P₃ were measured using the PI(3,4,5)P₃ mass strip kit. C, WT MEFs were treated with 3-MA (5 mm) or wortmannin (50 mm) for 1, 3, 6, and 9 h in full medium, and the intracellular level of PI(3,4,5)P₃ was measured using a chromatography-based method, as described under “Experimental Procedures.” The changes of PI(3,4,5)P₃ are shown as relative peak height and are presented as means ± S.D. from three independent experiments and analyzed using Student's t test (**, p < 0.01).