Figure 3. Assessment of autophagy regulated by 17β-E2 in osteoblasts using monodansylcadaverine (MDC) staining. The osteoblasts were pretreated with or without 3-MA for 2 h, and then cultured with 17β-E2 (0, 10-8, 10-6 M) for 24 h. Level of cell autophagy was evaluated by the intensity of fluorescent dots. The histogram showed that the relative intensity of fluorescent dots was significantly greater in 10-6 M 17β-E2 group than in the control group and 17β-E2 (10-6 M) + 3-MA group. Scale bar = 20 μm. The data were expressed as the means ± SD from 3 independent experiments. Statistically significance was evaluated using the Student’s t-test. * P < 0.05 vs. the control group was considered significant.