Caspase-3 activity was assessed using a colorimetric assay (ab39401, Abcam) according to the manufacturer's instructions. The activities of caspase-3, caspase-8 and caspase-9 were determined using a Multiplex Activity Assay kit (ab219915, Abcam) according to the manufacturer's instructions with some modifications. The protocol in the following link was followed: https://www.abcam.com/ps/products/219/ab219915/documents/Caspase-3-Caspase-8-and-Caspase-9-Multiplex-Activity-Assay-protocol-book-v1d-ab219915%20(website).pdf. The caspase-3, -8 and -9 substrates were at room temperature prior to the experiment. All controls and samples were assayed in triplicate. The cells were plated overnight in DMEM at 2×104/90 μl per well. Blank wells (DMEM without cells) contained 10 μl of compound buffer. To assay caspase-3 activity in each well, assay loading solution was prepared by the addition of 50 μl of caspase-3 substrate to 10 ml of Assay Buffer. To assay tri-caspase activity in the same well, a mixed assay solution was prepared by the addition of 50 μl of tri-caspase substrate to 10 ml of assay buffer together. This was followed by the addition of 100 μl/well of solution directly into cell plates without removing DMEM at room temperature for 45 min. A micro-plate reader (InfiniteM200, Tecan Group Ltd.) was used to measure fluorescence at Ex/Em=535/620 nm (caspase-3, red), Ex/Em=490/525 nm (caspase-8, green), Ex/Em=370/450 nm (caspase-8, blue).

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