The primary cultured β-cells were divided into 12-well plates and categorized into four groups. Each group was treated with different reagents as follows: (1) 5 mM STZ; (2) 5 mM STZ and 100 µM allantoin; (3) 5 mM STZ, 1 µM KU14R and 100 µM allantoin; and (4) control. The cells were incubated with the reagents for 48 h. Then, the cells were collected and the apoptotic cells in each group were quantified using Annexin V-PI staining (Life Technology, Carlsbad, California, USA) and analyzed using flow cytometer based on the previously described method (Luo et al., 2014 (link)).
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