Fig. 1.

Comparison of biguanide and diguanide compounds on cells. Structures of biguanides (a) and diguanides (b) are shown. (c, d) H4IIE liver cells had serum removed for 2 h and then were treated with the compounds shown for 3 h, prior to lysis and SDS-PAGE as detailed in the Methods. Immunoblotting was performed using two ACC antibodies, one that detects ACC phosphorylated on Ser79 (pACC Ser 79) and another that detects ACC whether or not it is phosphorylated. A third antibody detects AMPK when it is phosphorylated on Thr 172 (pAMPK Thr 172) and a fourth detects AMPK whether or not it is phosphorylated. Finally two antibodies that detect S6 protein were used. One detects S6 phosphorylation of residues 240 and 244 (pS6 240/244), whilst a second detects S6 regardless of phosphorylation. For each section, the phosphorylation responses shown are representative of observations from at least three similar experiments. Quantitation of replicates by densitometry appears in the supplementary Fig. 1.