Lung morphogenesis effects are AMPK dependent, whereas pancreatic cyst formation is AMPK independent. (A) Lkb1^WT/WT (WT) and Lkb1^MG/MG (MG) MEFs were seeded into 96-well plates with DMSO or 1 µM 1NMPP1 or 1 µM 1NMPP1 and 100 µM A-769662. At 72 h, cell nuclei were counted using Vybrant green DNA stain and ATP levels were measured using CellTitre-Glo. Error bars represent standard deviations (n = 12). (B) Mesenchyme-free Lkb1^WT/WT and Lkb1^MG/MG lung epithelium were cultured with 1 µM 1NMPP1 and 100 µM A-769662. The branching defect induced by 1NMPP1 in the mesenchyme-free Lkb1^MG/MG lung epithelium is reversed by the addition of A-769662. The images shown are E-cadherin–stained explants. Bar, 100 µm. (C) Pancreatic explants from E14.5 Lkb1^WT/WT and Lkb1^MG/MG were cultured with 1 µM 1NMPP1 and 100 µM A-769662. The cyst development induced by 1NMPP1 in the Lkb1^MG/MG explant is not reversed by the addition of A-769662. The images shown are maximum projections across an 18-µm z-stack of ZO-1–stained explants. Bars, 100 µm. (D) Embyronic explants (pancreas) from Lkb1^WT/WT (lanes 1, 2, 5, and 6) and Lkb1^MG/MG (lanes 3, 4, 7, and 8) embryos were cultured ex vivo on transwell filters and incubated 2 h with DMSO (lanes 1 and 3) or 1 µM 1NMPP1 (lanes 2, 4, 5, and 7) or 1 µM 1NMPP1 and 100 µM A-769662 (lanes 6 and 8) before collecting lysates. Western blots were analyzed with antibodies to Phospho-ACC(Ser79), Phospho-RAPTOR(Ser792), Phospo-AMPKα(Thr172), Lkb1, Phospho-S6(Ser235/236), and eIF4E (as loading control).