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. 2015 Sep 28;125(11):4063–4076. doi: 10.1172/JCI82152

Figure 2. LKB1 suppresses CCL2 production by human EM cells via an AMPK-dependent mechanism.

Figure 2

(A) Human CCL2 ELISA of conditioned media harvested 24 hours after plating of EM cells previously transduced with lentivirus. LKB1 knockdown led to a significant increase of CCL2 in the media (n = 3 biological replicates per experiment). (B) Human CCL2 ELISA on conditioned media containing AICAR (0.5 mM), metformin (5 mM), or vehicle (PBS) only. AICAR or metformin significantly reduced CCL2 secretion 24 hours after addition of drug (n = 3 biological replicates). (C) Representative Western blot of lysates from cells shown in panel B revealing partial restoration of pAMPK levels. (D) Human CCL2 ELISA on conditioned media harvested 24 hours after plating cells transduced with control or AMPKα1/2-shRNA lentivirus. AMPK knockdown led to a significant increase in CCL2 (n = 3 biological replicates per experiment). (E) Representative Western blot of lysates from cells shown in D showing knockdown of AMPK and undetectable pAMPK levels. *P < 0.05; **P < 0.001; ***P < 0.0001, Student’s t test. Error bars = SEM.