Journal: The Journal of Biological Chemistry

Article Title: Artesunate Activates Mitochondrial Apoptosis in Breast Cancer Cells via Iron-catalyzed Lysosomal Reactive Oxygen Species Production *

doi: 10.1074/jbc.M110.210047

Figure Lengend Snippet: ART disrupts localization of endolysosomes but not apparent function of critical lysosomal parameters. A, wild-type MCF-7 cells were subjected to the indicated experimental conditions for 24 h and then labeled with the lysosomal pH indicator LTR (5 nm) and analyzed by fluorescence microscopy. B, MCF-7 cells stably expressing GFP-Rab7 were incubated with MR cathepsin B substrate (2.2 μm) following 24 h of drug treatments. Both GFP-Rab7 and MR were analyzed by fluorescence microscopy. C, Rab7, LTR, and MR staining and intensity patterns were analyzed in MCF-7 wild-type cells (LTR) and cells stably expressing GFP-Rab7 (Rab7 and MR) following 3 h of treatment. D, flow cytometry was used to quantify population level responses. The mean fluorescence intensity of lysosomal LTR (0.1 μm) and MR (2.2 μm) was determined. Graphs indicate responses as the percentage of dye fluorescence normalized to KHS, and the x axis crosses at 100% (*, p < 0.05; #, p < 0.001, compared with KHS).

Article Snippet: LysoTracker Red, YO-PRO-1, propidium iodide, and H 2 DCF-DA were purchased from Invitrogen. ( z -RR) 2 -MagicRed-cathepsin B substrate was purchased from B-Bridge International Inc. TNF was a kind gift of BASF (Mannheim, Germany).

Techniques: Labeling, Fluorescence, Microscopy, Stable Transfection, Expressing, Incubation, Staining, Flow Cytometry