Betulinic Acid Induces ROS-Dependent Apoptosis and S-Phase Arrest by Inhibiting the NF-κB Pathway in Human Multiple Myeloma

Figure 4

BA induced ROS accumulation in U266 cells in a concentration-dependent manner. (a) Representative plots of flow cytometric analysis using a DCFH-DA probe after 12 h of treatment and the quantitative analysis of the intracellular ROS level. and . (b) Intracellular MDA content in different concentrations of BA; . (c) SOD activity in different concentrations of BA; . (d) Expression of SOD2, FHC, GCLM, and GSTM by real-time PCR after 40 μM BA for 12 h; . (e) Representative cytometric graphs and quantitative analysis of the ROS generation level with or without 10 mM NAC for 12 h. vs. the control group; ^##vs. the BA-treated group. (f) U266 cells were treated with 40 μM BA with or without 10 mM NAC, the apoptosis rate was detected by flow cytometry, and quantitative analysis was shown. vs. the control group; ^###vs. the BA-treated group. (g) Expression levels of apoptosis-related proteins were evaluated by Western blotting after treatment with or without 10 mM NAC.

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