Figure 5

Dapagliflozin rescued C2 ceramide-induced cell death in HK2 cells. (a) HK2 cells were incubated with 2 μM dapagliflozin for 36 h with or without 7.5 ng/ml C2 ceramide, and the expression levels of CHOP mRNA were determined by qRT-PCR (n = 5 independent experiments). (b) HK2 cells were treated with 2 μM dapagliflozin for 36 h with or without 7.5 ng/ml C2 ceramide, and caspase-3 activity levels were determined (n = 5 independent experiments). (c,d) HK2 cells were treated with 2 μM dapagliflozin for 48 h with or without 7.5 ng/ml C2 ceramide, and the cells were subjected to microscopic observation (c) and protein assay (d). These are representative images from experiments independently performed five times. (e), (f) HK2 cells were treated with 2 μM dapagliflozin for 48 h with or without 7.5 ng/ml C2 ceramide, and apoptosis and necrosis assay was performed using a Cell Death Detection ELISAplus (Roche, Indianapolis, IN).