Figure 2.

AMPK activators inhibit alkaline phosphatase activity during osteogenic differentiation. MC3T3‐E1 cells were incubated with osteogenic differentiation medium in the presence of different doses of metformin (Met) and Ibuprofen. After one week, the Alkaline phosphatase activity (metformin (Met) (A) and Ibuprofen(C)) was measured. The graph represents averages of three independent experiments (mean ± SD, N = 3). *Significant difference from metformin (A) and Ibuprofen(C) versus control was assessed using Student's t test (*P < .05). Western blot (metformin (B) and Ibuprofen(D)) was also performed to study activation of AMPK and Smad1/5. NM: no differentiation medium, α‐MEM complete medium