Figure 2. AMPK activation induces PGC-1α mRNA expression and transcriptionally activates the PGC-1α promoter.

C₂C₁₂ cells were treated with either AICAR (1 mM) or Vehicle for 24 hrs. A. Representative Western Blot probed with a Phospho-AMPKα (Thr172), stripped and then re-probed with total AMPKα for loading control (upper panel). Summary of repeated experiments of the effect of AICAR on AMPK activation is shown (lower panel; n = 4). B. upper panel, EtBr-stained DNA gel of PGC-1α amplified by PCR from vehicle- and AICAR-treated cells. GAPDH was also amplified by PCR and used to verify equal loading. Lower panel: A summary of repeated experiments of the effects of AICAR on PGC-1α mRNA expression (n = 3). C. AICAR-induced transcriptional regulation of the PGC-1α promoter. Relative luciferase activity of the PGC-1α promoter constructs in vehicle- or AICAR-treated cells is shown (n = 4–6). D. The AICAR-responsive region (ARR) from −473 to −821 was cloned into the pGL4.23 minimal promoter vector and AICAR-induced transcriptional regulation of this region was assessed (n = 3). For all data, values are means±S.E.M, *, p<0.05 versus Vehicle-treated control; §, p<005 versus pGL4.23.