Fig. 6. Overexpression of GPx1 in PDA cells in glucose-free medium promotes cell death.

a PANC-1 cells ectopically overexpressing GPx1 were established. b The expression levels of LC3, SQSTM1 and β-actin were assessed by immunoblot analysis in PANC-1 cells with or without GPx1 overexpression in DMEM or glucose-free medium for 24 h. c, d ROS and H₂O₂ levels were measured in PANC-1 cells with GPx1 overexpression (**P < 0.01). e, f PANC-1 cells were treated with or without NAC (20 mM) in DMEM or glucose-free medium for 24 h. Apoptosis was analyzed by using the annexin V/7-AAD assay (**P < 0.01, compared with all other groups by Tukey’s post hoc test). g, h Apoptosis was analyzed in PANC-1 cells with or without GPx1 overexpression in DMEM or glucose-free medium for 24 h (**P < 0.01). i PANC-1 cells were treated with or without NAC (20 mM) in DMEM or glucose-free medium for 24 h. Cell lysates were immunoblotted for cleaved PARP, cleaved caspase-3, and β-actin. j Expression levels of cleaved PARP, cleaved caspase-3, and β-actin were assessed by immunoblot analysis in PANC-1 cells with or without GPx1 overexpression cultured in DMEM or glucose-free medium for 24 h