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After all time points were collected, fixed cells were pelleted and washed with DPBS twice, then resuspended in 250 l of 50 g ml 1 propidium iodide (Biotium, 40017) with 100 g ml 1 RNase A (QIAGEN) staining solution for 1 h at room temperature or overnight at 4 C, protected from light

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After Harleys death in 2000, a club was formed to preserve his legendary accomplishments

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This can take 30 to 90 minutes before effects begin, depending on your metabolism, body weight, and whether youve eaten recently