Figure 2

Dapagliflozin regulates unfolded protein response in HK-2 cells. (a) HK2 cells were incubated with 2 μM dapagliflozin for 36 h and collected for protein expression analysis. These immunoblots are representative of n = 5 experiments performed independently. (b) Signal quantification of the expression levels for S⁵¹ phosphorylation of elf2α normalised with total elf2α levels. (c) HK2 cells were incubated with 2 μM dapagliflozin for 36 h, and the ATF4 mRNA expression levels were determined by qRT-PCR (n = 5 independent experiments). (d) HK2 cells were incubated with 2 μM dapagliflozin for 36 h, collected for protein analysis, and immunoblotted for the indicated proteins. These are representative immunoblots independently performed five times. (e,f) Signal quantification of the expression levels of ATF6 normalised with tubulin-α (e) and the expression levels for S⁷²⁴ phosphorylation of IRE1α normalised with total IRE1α levels (f). (g) HK2 cells were incubated with 2 μM dapagliflozin for 36 h, and the expression level of CHOP mRNA was determined by qRT-PCR (n = 5 independent experiments). (h) HK2 cells were incubated with 2 μM dapagliflozin for 36 h and caspase-3 activity levels were measured using a caspase-3 fluorescence assay kit (MBL) (n = 5 independent experiments).