Enhancement of the Acrolein-Induced Production of Reactive Oxygen Species and Lung Injury by GADD34

Figure 5

GADD34 promote recovery from a shutoff of total protein synthesis and enhance cell death. The 25 μM acrolein-treated shcon/LLCs and shGADD34/LLCs were analyzed. (a) Cell survival after 25 μM acrolein treatment was measured. . (b) Levels of ROS production in shcon/LLCs and shGADD34/LLCs were measured by DCFH-DA after 25 μM acrolein treated for 8 h. (c) Cells were collected at the indicated times and protein expressions of ER stress signaling were detected by western blot. (d) The 25 μM acrolein-treated shcon/LLCs and shGADD34/LLCs with 20 mM NAC or without NAC for 8 h. Cells were stained with Annexin V-PE/7-amino-actinomycin D (7AAD). (e) Bands of proteins were analyzed by SDS-PAGE after 25 μM acrolein treatment. (f) Protein concentration was measured by Bio-Rad Protein Assay. (g) The shcon/LLCs and shGADD34/LLCs were treated by 25 μM acrolein with 10 μM MG132. Cells were collected and lysate was analyzed by SDS-PAGE. (h) Amount of proteins was measured by Bio-Rad Protein Assay. Data are represented as means ± s.e.m. .

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