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. 2018 Sep 18;9:1315. doi: 10.3389/fphys.2018.01315

FIGURE 5.

FIGURE 5

Lysophosphatidic acid down-regulates autophagy through the AKT/mTOR pathway. (A,B) Western blotting analysis of the phosphorylation of AMPKα (Thr172), mTOR (Ser2448), P70S6 kinase (Thr389), and 4E-BP1 (Thr37/46) in H9C2 cardiomyoblasts treated with 10 μM LPA for the indicated time. ∗P < 0.05 vs. control, nsP > 0.05. (C) H9C2 cardiomyoblasts were treated with 50 nM rapamycin for 1 h, before 10 μM LPA stimulation for 15 min. Then the phosphorylation of mTOR and P70S6 kinase were analyzed by Western blotting. ∗P < 0.05, ∗∗P < 0.01. (D) H9C2 cardiomyoblasts were treated with 6.25 μM LY294002 for 1 h, and then exposed to 10 μM LPA for 15 min. The phosphorylation of mTOR and p70S6 kinase were analyzed by Western blotting. ∗P < 0.05, ∗∗P < 0.01. (E) H9C2 cardiomyoblasts were treated with 6.25 μM LY294002 for 1 h, before exposure to 10 μM LPA for 4 h. Then the protein levels of LC3B were determined by Western blotting. ∗P < 0.05. All data represents mean ± SEM. Each experiment was performed thrice and shown results were representative. A one-way ANOVA followed by a post hoc Tukey’s test was used for multiple comparison.