Depletion of Luminal Pyridine Nucleotides in the Endoplasmic Reticulum Activates Autophagy with the Involvement of mTOR Pathway
Figure 4
Time course of the effect of high concentration metyrapone on ER stress. HepG2 cells were treated with 100 μM metyrapone for 5–120 min at 37°C. (a) Cell lysates were analysed by Western blot by immunoreacting with antibodies against LC3, PARP, and GAPDH, respectively. The relative density of both the lower band of LC3/GAPDH and PARP/GAPDH were represented. (b) Cell lysates were analysed by Western blot by immunoreacting with antibodies against ER stress markers, such as eIF2α-P, eIF2α, CHOP/Gadd153, and Gadd34, respectively. The relative density of Gadd34/GAPDH was represented. (c) Cell viability was assessed by counting the cells both permeable and non-permeable to trypan blue. About 5% of the cells were nonpermeable for trypan blue.
