Fig. 2.

Anaplerotic factors pyruvate or oxaloacetate, similar to αKG, prevent AOA-induced effects on cell proliferation, mTORC1 and mTORC2 activity in WI38 cells. WI38 cells were treated with vehicle or AOA in the absence or presence of 3 mM pyruvate, oxaloacetate or 5 mM αKG for the indicated time-period with medium changed at 2-day intervals. (A) Effect of anaplerotic factors on AOA-induced inhibition of cell proliferation. Cell numbers were assessed and calculated as described in Fig. 1A. (B) Effect of anaplerotic factors on AOA modulation of mTORC signaling (n=3). After 24-h treatment, cell lysates were prepared and analyzed for the activation status of mTORC1 and mTORC2 respectively indicated by P-S6K1-T389/S6K1 and P-AKT-S473/AKT as described in Fig. 1B. All quantitative data are expressed as the mean±s.e.m. (n=3) of three independent experiments. Different lowercase letters indicate significant difference among treatment groups at the same time-point (P<0.05). * and # designate a significant difference compared with the respective vehicle control and AOA group, respectively (P<0.05).