Caspase glo 3 7 and 9 assay kits

Caspase 3 and 9 catalytic activities were assessed using Caspase-Glo 3/7 and 9 assay kits (Promega, USA) according to the manufacturers’ instructions (16 (link), 32 (link)). For Caspase-Glo 9 substrates, the mixture was treated with the inhibitor MG132. Briefly, HCT116 cells were seeded at a density of 2×10³ cells/well in a white-walled 96-well plate and cultured for 8 h. HCT116 cells were subsequently incubated with or without 54.3 μM GA-Me for 24 hours. Then, 100 μl of medium were carefully removed from each well without disturbing the cells before 100 μl of Caspase-Glo reagent were added to the cells. The plate was placed on an orbital shaker for 30 s and subsequently incubated at room temperature for 1 hour. The luminescence was measured using a GENios Pro plate reader (Tecan, Research Triangle Park, NC, USA).