FIGURE 5. Metformin inhibited adipocyte mediated lipid accumulation in ID8 cells.

(A) ID8 cells were grown in 50% adipocyte CM (adipo CM) or CM from adipocytes differentiated in presence of metformin 2 or 4 mM (adipo M4 CM, or adipo CM) in the presence of metformin (adipo CM + M4) for 72-96 hours in chamber slides. The cells were stained with Bodipy 493/503 and counterstained with Hoechst 33342 as described in the Methods. The lipid droplets were stained green and visualized with a fluroscent microscope at 40x magnification. (B) Under same conditions, the protein extracted was subjected to western blot and immuno-blotted for expression of phosphorylated (p) and total (t) adenosine monophosphate activated kinase (AMPK) and acetyl CoA carboxylase (ACC). Micrographs and blots are representative of 2 individually performed experiments.