Figure 8

Effect of NAC on ROS content, cell death , lysosomal membrane permeability and the leakage of CatB to the cytosol.

To identify the effect of NAC in vitro, cells were treated with 100 μg/ml of NAC for 12 h followed by an additional 2 h with 500 μM H₂O₂ for the assay with FACS. For the ROS content assay, the cells were treated with 100 μg/ml NAC for 24 h followed by 500 μM H₂O₂ for different amounts of time. In other assays, the cells were treated with 100 μg/ml NAC for 16 h followed by an additional 8 h with 500 μM H₂O₂. (A) The Annexin V/PI assay with FACS is shown. (B) The ROS contents in cells is shown. (C) LysoTracker Green staining (scale bar = 10 μm) is shown. (D) Quantification of the LysoTracker Green staining is shown. (E,H) Levels of LAMP1 protein were measured in cells by Western blotting and quantification is shown. (F,G,I,J) Cat B/D levels were measured in the cytosol/whole lysate by Western blotting and the quantification is shown. (*p < 0.05, **p < 0.01).