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. Author manuscript; available in PMC: 2015 May 16.
Published in final edited form as: ACS Chem Biol. 2015 Feb 17;10(5):1258–1268. doi: 10.1021/cb500772c

Figure 4.

Figure 4

PLD inhibitor treatment decreases mTOR signaling events. (A, B) Phosphorylation at serine 65 phosphosite on 4E-BP1 after treatment of U87MG cells (A) and U87MG (Myr-Akt) (B) with different inhibitors. Treatment with 10 μM VU0359595 decreased phosphorylation at this site, validating the findings of the SILAC phosphoproteomics screen. (C, D) Magnitude of decrease in phosphorylation was similar in U87 parental (C) and U87Myr-Akt (D) cells, confirming the Akt independence of the changes in mTOR signaling induced by the PLD inhibitor. (E, F) siRNA knockdown of the PLD enzymes confirmed that PLD1 knockdown specifically decreased phosphorylation of serine 65 on 4E-BP1. Statistics evaluated by ANOVA; *p < 0.05; **p < 0.01.