Caspase-Glo 3/7 Assay

About 10000 cells/well of UMUC-5, 5637, and UMUC-3 cell lines were incubated overnight in white walled, clear bottom 96-well plates. The apoptosis inducer, Staurosporine (1µM) (SelleckChem) (PubChem CID – 44259), was incubated with the cells for 3–4 hours in the presence and absence of the caspase inhibitor Z-VAD-FMK (20µM) (Promega Corporation) (PubChem CID – 5497174). Experimental wells were treated with 10µg/ml of pan/pan IR700 or an equivalent concentration of IR700 with or without Z-VAD-FMK (20µM) for 1 hour followed by irradiation with an appropriate amount of NIR (UMUC-5 – 4 J/cm², 5637 – 32 J/cm² and UMUC-3 – 64 J/cm²). Approximately 20 to 30 minutes post-NIR treatment, 100µl of Caspase-Glo 3/7 reagent was added to each well. Plates were incubated at room temperature for 30 minutes and luminescence was measured using EnSpire multimode plate reader (Perkin Elmer).