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. 2014 Sep 12;141(1):244–253. doi: 10.1093/toxsci/kfu122

FIG. 1.

FIG. 1.

Cytotoxicity and intracellular generation of ROS in CsA- and FK506-treated HepaRG cells. (A) Cells were incubated for different time points (0–24 h) with different concentrations of either CsA or FK506 (0–100μM). Cytotoxicity was measured by the MTT colorimetric assay. (B) Cells were treated with different concentrations of either CsA or FK506, or co-treated with 50μM CsA + 15mM NAC or 5mM PBA for 2 h. H2O2 at 25mM was used as a positive control. ROS generation was detected by the DCFDA specific substrate. (C and D) Cells were treated with CsA or FK506 or co-treated with 50μM CsA + (15mM NAC or 5mM PBA) for 6 h, then mRNA levels of ROS markers (HO-1, MnSOD, and Nrf2) and ER stress markers (CHOP, GRP78, ATF4, and ATF6) were estimated by RT-qPCR. Data represent the means ± SD of three independent experiments. All results are expressed relative to the levels found in untreated cells, arbitrarily set at 1 or 100% *p < 0.05, **p < 0.01, ***p < 0.001 compared with non-treated cells, #p < 0.05, ##p < 0.01, ###p < 0.001 compared with 50μM CsA.