Fig. 2. Nerolidol induced G1 in Nerolidol inhibits proliferation of leiomyoma cells via reactive oxygen species-induced DNA damage and downregulation of the ATM/ Akt pathway

Fig. 2. Nerolidol induced G1 cell cycle arrest by downregulating Akt phosphorylation and cell cycle-related proteins. ELT3 cells were treated with the indicated concentrations of nerolidol for 48 h and were then harvested after trypsin/EDTA treatment. (A, B) For cell cycle analysis, the harvested cells were fixed with 70 % ethanol, followed by propidium iodide staining, and the DNA content was measured using flow cytometry. The results were analyzed using the FlowJo software (A). Blue, olive green, and green represent the G1, S, and G2/M phases, respectively. The percentage of each cell cycle phase was plotted in (B). The p-value was calculated using one-way ANOVA with Tukey's multiple post hoc test for each group.*, p <0.05; **, p <0.01, and ***, p <0.001 compared to the vehicle control group (1 % EtOH). (C) Total cellular protein content was measured, and the expression levels of phosphorylated Akt, total Akt, cyclin D1, CDK4, and CDK6 were determined using western blot analysis. β-Actin was used as the loading control. The inserted numbers in (C) indicate the relative expression level compared to that of the vehicle control (indicated as 0 μM). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)