Figure 2.

MK-2206 treatment increases ROS in primary and immortalized leiomyoma cells. A, Primary leiomyoma cells treated with vehicle (DMSO), BEZ235 (phosphatidylinositol 3-kinase/mTOR inhibitor) or MK-2206 (AKT inhibitor). Immunofluorescent staining of ROS, using DHE was done (left), and the relative levels of ROS were quantitatively measured (right). B, Primary leiomyoma cells were treated with H₂O₂ (100 μM) for 8 hours or 24 hours. Number of β-gal-positive staining cells was counted. C, Leiomyoma cell lines were treated with H₂O₂ (100 μM), and expression of senescence-associated genes GLB1, P16, and P53 was measured by real time RT-PCR. D, Leiomyoma cell lines were treated with MK-2206 (2 μM), MK-2206 (2 μM) with NAC, and H₂O₂ (100 μM), and expression of P16 and P53 was measured by real time RT-PCR. *, P < .05; **, P < .01; ***, P < .001.