Figure 2
Res-006 induces cell death associated with ER stress and/or ROS in HepG2 cells. (A) Flow cytometry analysis of cell death after Res-006 (65 μmol/L) treatment for 24 h. Representative images are shown (n=3 per group). (B) HepG2 cells were treated with Res-006 (65 μmol/L), and total cellular extracts were prepared and subjected to Western blot analysis. (C) HepG2 cells were treated with Res-006 (65 μmol/L), Res-006 (65 μmol/L), and NAC (1 mmol/L), or with Res-006 (65 μmol/L) and 4-PBA (0.5 mmol/L) for 24 h, and the cell viability was analyzed by the CCK-8 assay. The data are expressed as the mean±SEM of three independent experiments, **P<0.01 and ***P<0.001; untreated group vs each treated group, ##P<0.01 and ###P<0.001; Res-006-treated group vs each treated group. (D) HepG2 cells were cotreated with Res-006 (65 μmol/L) and BAPTA/AM (0–5 μmol/L) for 24 h, and cell viability was analyzed using the CCK-8 assay. The data are expressed as the mean±SEM of three independent experiments, ***P<0.001; untreated group vs each treated group. (E) HepG2 cells were transfected with pCMV-Flag3x-ATF6α for 48 h and then treated with Res-006 (65 μmol/L) for the indicated times or were mock treated, with Tm (10 μg/mL) and Tg (1 μmol/L) for 12 h, and total cellular extracts were prepared and subjected to Western blot analysis.
