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. 2020 Mar 25;25(7):1479. doi: 10.3390/molecules25071479

Figure 5.

Figure 5

Analysis of typical features of intrinsic apoptosis pathway in HL-60 cells treated with DL-247 for 24 h. (A) Percentage of phosphorylated H2AX positive cells measured by flow cytometry (Alexa Fluor 647 Mouse Anti-H2AX (pS139) antibody staining). (B) Mitochondrial membrane potential (ΔΨm) changes measured by flow cytometry (JC-1 staining). FCCP (30 μM, 30 min incubation at 37 °C) was used as a positive control. (C) Bax and Bcl-2 gene expression changes analyzed by real-time PCR. (D) Intracellular ROS generation measured by flow cytometry (CellROX Green Reagent staining). Menadione (200 µM, 1 h incubation at 37 °C) was used as a positive control. (A–D): Data are presented as mean ± SEM of three independent experiments. (A,B,D): Statistical significance was assessed using one-way ANOVA and a post-hoc multiple comparison Student–Newman–Keuls test. * p < 0.05, *** p < 0.001 vs. control. C: Statistical significance was assessed using Student-t test. * p < 0.05, *** p < 0.001 vs. control.