Fig. 7

ISO decreases the level of ROS during RANKL-induced osteoclastogenesis by enhancing the expression and nuclear translocation of Nrf2 and increasing the expression of antioxidant enzymes. A RAW264.7 cells were stimulated with RANKL in the absence or presence of the indicated concentrations of ISO. The intracellular ROS levels were assessed by quantifying the intensity of the fluorescence of DCF, and representative confocal images were described. B Quantification of the average DCF fluorescence intensity of the cells in each well. C RAW264.7 cells were pretreated with or without 100 μM ISO and then stimulated with RANKL; Nrf2 immunofluorescence staining (red), and nuclei (blue) were labeled with DAPI. D Representative western blot images showing the expression of Nrf2 and several antioxidant enzymes (CAT, GCLC, and HO-1), normalized to β-actin. RAW264.7 cells, stimulated with RANKL in the absence or presence of ISO (0, 20, 40, 60, 100 μmol) for 2 d, were used for protein extraction and analysis. E–H Quantitative analysis, the band intensity of each protein was normalized to that of β-actin. Values were presented as mean ± standard deviation (n = 3); Scale bar = 100 μm; *p < 0.05, **p < 0.01