Research Paper Volume 4, Issue 7 pp 480—498

Figure 4. Effects of the folate-independent salvage pathway of nucleotide biosynthesis on metformin-induced activation of AMPK. Left. Western blot analysis of total and phosphorylated AMPK (fosfo-AMPKα^Thr172). Equal amounts of total protein (50 μg) were loaded in each line; use of β-actin as a control showed equal loading between lanes. Metformin (MET) was used at 1 mmol/L, thymidine was at 5.6 μmol/L, and hypoxanthine was at 32 μmol/L; drug exposure was 48 h. Right. Immunofluorescence for phospho-AMPKα^Thr172 in MDA-MB-231 cells either untreated or treated with 1 mmol/L metformin (MET) in the absence or presence of thymidine plus hypoxanthine for 48 h. Images were captured in different channels for phospho-AMPKα^Thr172 (green) and Hoechst 33258 (blue) with a 20x objective. (MET, metformin).