Figure 4: Continuous high (25 mM) glucose induces apoptosis of BRP. Subconfluent cells in 30 mm2 dishes were exposed to normal (N, 5.6 mM) and high (CG, 25 mM) glucose for 4 days. (a) High glucose activates caspase-3. After 4 days, the cells were washed twice with ice-cold PBS, lysed and apoptosis was measured using colorimetric caspase activity assay. Data are represented as Means ± SEM of 6 separate experiments. P*=.0315 versus N. (b) High glucose induces DNA fragmentation. After 4 days, DNA fragmentation was measured using Cell Death Detection ELISA plus (Roche, Hertfordshire, UK). Data are represented as Means ± SEM of 5 separate experiments. P=.043 versus N.