Fig 1: Metformin inhibits phosphorylation of ß-cateninSer552in ATDC5 cells. ATDC5 cells were transfected with AKT2 or treated with metformin (0.5, 1, 2, and 4 ?mM) and incubated for 2 (A–D), 4 (E–H), 6 (I–L), 12 (M–P) hours. At the end of the experiment, cell lysates were collected and Western blot analysis was performed (n ?= ?3). Metformin significantly inhibits ß-cateninS552 phosphorylation, cyclin D1 and Axin2 expression. Cyclin D1 and Axin2 are downstream target genes of ß-catenin. Metformin inhibited ß-cateninS552 phosphorylation but had no significant effect on total ß-catenin levels. The data are expressed as mean ?± ?SD, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 and ns, not significant.