The inhibitory effects of PSH on cell proliferation were measured by flow cytometry. Following the aforementioned treatments, U251 cells were collected and fixed with 70% ethanol at 4°C for 2 h. Following centrifugation (800 × g for 5 min at 4°C), the cells were resuspended in RNase A solution and 0.02 mg/ml PI (Sigma-Aldrich; Merck KGaA), and incubated for 30 min at 4°C. The fluorescence of 1×104 cells was recorded by a FACSCalibur C6 flow cytometer (BD Biosciences) and analyzed by ModFit software (Verity Software House).

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