FOXO3 coordinates LKB1 gene expression depending on the metabolic status of the cell.

<p>MCF-10A cells were treated with AMPK activating stimuli (1 mM AICAR, 50 mM 2- Deoxyglucose or 2 µM Oligomycin) in combination either with vehicle (Ctrl), 1 µM dexamethasone (DEX) or a mixture of 1 µM dexamethasone and 1 µM RU-486 (DEX/RU) for 18 hours. (<b>A</b>) Relative mRNA levels of SGK-1, LKB1, FOXO3 and GAPDH as internal control were analysed by semi-quantitative RT-PCR and separated in a 1% agarose gel after ethidium bromide staining. (<b>B</b>) Relative mRNA levels of LKB1 and FOXO3 were analysed by quantitative real time PCR and normalized to GAPDH (mean ±SD, n = 3). (<b>C</b>) Western blot analyses of the relative protein levels of phospho-AMPK (T172), phospho-FOXO3 (S413), phospho-FOXO3 (T32), total FOXO3 in comparison to actin as internal control. The ratios of p-FOXO3/FOXO3 for each experimental condition are indicated. The data shown is representative of three independent experiments. (<b>D</b>) Schematic illustration underlying the glucocorticoid-dependent modulation of FOXO3 transcriptional activity.