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INS-1 832/3 cells were exposed overnight to thapsigargin (1 μM) in serum-free RPMI plus 11 mM glucose. Apoptosis was determined using Caspase Glo 3/7 (Promega). Signal was expressed relative to thapsigargin-only wells.
MIN6B1 cells were incubated overnight with serum-free media plus 25 mM glucose and 0.5 mM palmitate/BSA ± 100 nM agonists, fixed, processed with the In Situ Cell Death Detection Kit, TMR red (Roche), and imaged as above. Five images were taken in random areas and a minimum of 500 cells counted per experiment.
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