AMPK is an upstream positive regulator of ERK along AIA pathway of Jurkat T cells.

<p>(A) Jurkat T cells treated with the indicated periods of ANE 30–100K (30–100K) (9 μg/ml) or with the indicated concentrations of 30–100K for 1 hour were subjected to immunoblotting with phospho-ERK1/2 (p-ERK), total-ERK1/2 (t-ERK), and β-actin antibodies. (B) Cells treated with the indicated periods of 30–100K (9 μg/ml) in the presence or absence of U0126 (10 μM) were subjected to immunoblotting with phospho-AMPKα-Thr^<b>172</b> (p-AMPK), total AMPK (t-AMPK), p-ERK, t-ERK, and β-actin antibodies. (C) The same experiment was performed as (B), except that U0126 was substituted by compound C (Com C, 5 μM). (D) Lysates of parental (Pa), virus control (VC-A1), and sh-AMPK 3’UTR-A1 cells treated with the indicated periods of 30–100K (9 μg/ml) were subjected to immunoblotting with p-AMPK, t-AMPK, p-ERK, t-ERK, and xx-actin antibodies. (E) The same 30–100K treatment and immunoblotting were performed in sh-AMPK 3’UTR-A1-1 and 3’UTR-A1-2 cells and presented as (D). (F) Lysates of Jurkat T and OECM-1 cells treated with 30–100K (9 and 15 μg/ml, respectively) for the indicated periods in the presence or absence of U0126 (10 μM) were immunoblotted as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128011#pone.0128011.g001" target="_blank">Fig 1A</a>. Average LC3-II/actin ratios ± SD of 30–100K treatment for 24 hours of both cells were plotted. *<i>P</i> < 0.05, **<i>P</i> < 0.01.