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Figure 8 | Journal of Biomedical Science

Figure 8

From: Insulin receptor substrate-1 prevents autophagy-dependent cell death caused by oxidative stress in mouse NIH/3T3 cells

Figure 8

Oxidative stress induces autophagy-dependent cell death. (A) Two stable ATG-5 knockdown cell lines were examined for the presence of ATG-5 by Western blotting (left). The levels of ATG-5 and LC3B-II were normalized to that of actin for comparison over four independent experiments (right). (B) Control cells and the ATG-5 knockdown cells were treated with DMSO or GO for 6 h. ATG-5, LC3B-I, and LC3B-II levels were examined by Western blotting (left). The levels of LC3B-II were normalized to that of actin for comparison, using three independent experiments (right). (C) Control cells and the ATG-5 knockdown cells were treated with 10 mU/ml GO for 6 h. Cell death was examined using trypan blue dye exclusion assay. Three dishes of cells of each condition were observed for statistical analysis. The results for a representative sample of three independent experiments are shown. (D) Control cells and the ATG-5 knockdown cells were treated with 10 mU/ml GO for 6 h. Cells were collected using trypsinization and stained with PI. Cell death was analyzed from the proportional area of sub-G1 group DNA, as determined by flow cytometry. Eight independent experiments were performed for statistical analysis. M1 indicates the sub-G1 area.

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