Xihuang Pill Induces Apoptosis of Human Glioblastoma U-87 MG Cells via Targeting ROS-Mediated Akt/mTOR/FOXO1 Pathway

Figure 4

XHP-induced apoptosis in U-87 MG cells. (a) Confocal images of U-87 MG cells treated with different concentrations of XHP determined by TUNEL assay stained by FITC-dUTP and DAPI (× 200). (b) Effects of XHP on cell apoptosis in U-87 MG cells after 24h treatment determined by FACS following AnnexinV-FITC and propidium iodide double staining. The fluorescence of FITC and PI were measured in the FL1 channel and FL2 channel, respectively. A total of 10⁵ events were recorded for each sample before any gate setting and were analyzed with the FlowJo software V7.6. Cells in the right lower quadrant are undergoing early apoptosis. Cells in the right upper quadrant are undergoing late apoptosis. PTX was used as a positive control. Representative results are shown (n = 3). (c) U-87 MG cells were incubated with different concentrations of XHP, 2 µM PTX, or control medium for 24h, stained with AnnexinV-FITC and propidium iodide and subjected to flow cytometry analysis. Quantified data of apoptosis cells after treatments with varying formulations (n = 3, mean ± SD). (d) After being incubated with different doses of XHP, 2 µM PTX, or control medium for 24h, U-87 MG cells were harvested and subjected to western blot analysis with anti-Bcl-xL, anti-Bax, anti-caspase-3, and anti-caspase-9 antibodies.

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