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. 2010 May 19;285(29):22291–22298. doi: 10.1074/jbc.M110.123794

FIGURE 4.

FIGURE 4.

Transduction with siRNA targeting LKB1 diminished an AMPK signaling in HUVECs. A, LKB1 is required for full AMPK signaling. Western blot analysis for LKB1, phosphorylated ACC at Ser-79 (pACC), phosphorylated AMPK at Thr-172 (pAMPK), total AMPK (AMPK), and α-tubulin (Tubulin) is shown. Right panel shows quantification of LKB1, pAMPK, and pACC. *, p < 0.05 versus unrelated siRNA (n = 3 in each group). HUVECs were treated with siRNA targeting LKB1 for 48 h. B, LKB1 is required for metformin-stimulated eNOS phosphorylation. Western blot analysis for LKB1, phosphorylated ACC, phosphorylated eNOS (P-eNOS), and α-tubulin is shown. HUVECs were treated with siRNA targeting LKB1 for 48 h followed by treatment with metformin (2 mm) or vehicle for 3 h. A representative Western immunoblot is shown. C, LKB1 overexpression activates AMPK signaling and leads to an increase in eNOS phosphorylation. Western blot analysis for LKB1, phosphorylated-ACC at Ser-79, phosphorylated-AMPK at Thr-172, total AMPK, phosphorylated-eNOS at Ser-1177, total eNOS, and α-tubulin is shown. HUVECs were infected with Ad-β-gal or Ad-LKB1 for 24 h. Right panels show quantitative Western blot analyses for P-AMPK and P-eNOS. **, p < 0.01 versus Ad-β-gal treatment (n = 3 in each group).