To investigate cell cycle progression, harvested cells were stained with propidium iodide using the CycleTest Plus DNA Reagent kit (Becton–Dickinson, San Jose, CA, USA) according to the manufacturer’s recommendations. Flow cytometry was performed using a FACSCalibur flow cytometer (Becton–Dickinson) and the DNA histogram was analyzed using CELLQuest version 6.0 (Becton–Dickinson) [7 (link)].
Senescence was measured by the SA-β gal staining kit (Millipore, Billerica, MA, USA) according to the manufacturer’s protocol. In brief, cells were plated at a low density of 2,000 cells in each well of 12-well plates, and treated with the indicated concentrations of PAL or ABE for 3 days. Cells were then washed with PBS, fixed and stained with the SA-β gal solution (Millipore) for 4 hours or overnight. Senescent cells were quantified by counting 100 cells in 3 different fields for each replicate [7 (link)].
Senescence was measured by the SA-β gal staining kit (Millipore, Billerica, MA, USA) according to the manufacturer’s protocol. In brief, cells were plated at a low density of 2,000 cells in each well of 12-well plates, and treated with the indicated concentrations of PAL or ABE for 3 days. Cells were then washed with PBS, fixed and stained with the SA-β gal solution (Millipore) for 4 hours or overnight. Senescent cells were quantified by counting 100 cells in 3 different fields for each replicate [7 (link)].