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. 2015 Nov 10;157(1):268–281. doi: 10.1210/en.2015-1690

Figure 1.

Figure 1.

Biphasic regulation of AMPK activation is required for osteoblast differentiation. A, Cell lysates from MC-3T3 cells following exposure to DM for the indicated number of days were prepared and immunoblotted with an anti-pAMPK (T172) antibody as described in Materials and Methods. β-actin was immunoblotted as a loading control. Each bar is the ratio of the scan value of the pT172AMPK band divided by the AMPK band. B and D, MC-3T3 cells were stained by Alizarin Red on day 21 after DM alone (control) or DM plus compound C (1uM) (B) or metformin (0.5mM) (D), which had been added on the indicated day. C and E, Cell lysates were obtained from MC-3T3 cells on day 21 following DM exposure in the absence or presence of compound C (1uM) (C) or metformin (0.5mM) (E) that had been added on the indicated day. They were immunoblotted with antiosteocalcin or anti-β-actin. Each bar is the ratio of the scan value of the osteocalcin band divided by the β-actin band. **, P < .01 and ***, P < .001 indicate significant differences between two treatments.