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. 2016 Jan 18;2:15072. doi: 10.1038/cddiscovery.2015.72

Figure 1.

Figure 1

Effect of Imeglimin on tBH- or hyperglycemia-induced endothelial cell death. HMEC-1 cells incubated in the absence or presence of either 100 μM Imeglimin for 24 h, 10 mM Imeglimin for 4 h or 1 μM CsA for 30 min were washed with PBS before subsequent exposure to 0.5 mM tBH or vehicle (control) in FBS-free culture medium for 45 min. Cells were then washed with PBS and incubated at 37 °C for 24 h in a complete MCDB medium. Alternatively, HMEC-1 cells were cultured for 48 h in a complete MCDB medium at the indicated concentration of glucose, in the absence or presence of 1 μM CsA, 250 μM N-acetyl cysteine (NAC) or 100 μM imeglimin. Cytotoxicity was evaluated by staining cells with Alexa Fluor-conjugated annexin V and PI. (a and b) Representative data. (c and d) Percentage of dead cells (i.e., cells positive for annexin V or PI) in five different experiments. Results are mean±S.E.M.; # P<0.01, *P<0.05 versus control cells, paired Student’s t-test.