Biomolecules & Therapeutics : eISSN 2005-4483 / pISSN 1976-9148

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Fig. 5. BITC reduced the lysosomal cathepsin activity and caused lysosomal swelling in AGS cells. (A) AGS cells were treated with 5, 10, or 15 µM BITC for 24 h. Whole cell lysis was performed, and the supernatant was incubated with omnicathepsin (100 µM) for 30 min at 37°C. The emitted fluorescence was quantified in a plate reader (exc: 380 nm, em: 440 nm) and normalized by total protein concentration. (B) AGS cells were treated with 10 µM BITC for 6, 12, or 24 h. Whole cell lysis was performed, and the supernatant was incubated with omnicathepsin (100 µM) for 30 min at 37°C. The emitted fluorescence was quantified in a plate reader (exc: 380 nm, em: 440 nm) and normalized by total protein concentration. (C) AGS cells were treated with or without 100 nM bafilomycin A1 for 24 h. Whole cell lysis was performed, and the supernatant was incubated with omnicathepsin (100 µM) for 30 min at 37°C. The emitted fluorescence was quantified in a plate reader (exc: 380 nm, em: 440 nm) and normalized by total protein concentration. (D) AGS cells were treated with 5, 10, or 15 µM BITC for 24 h, stained with 1 µg/ml AO for 20 min at room temperature, and visualized by a confocal microscope within 1 h (scale-bar=50 μm). AO emits red fluorescence in the acidic compartment and green fluorescence in the cytoplasm. Data were calculated from three independent experiments and expressed as the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 versus the control, and #p<0.05, ###p<0.001 versus BITC treatment alone.
Biomolecules & Therapeutics 2022;30:348~359 https://doi.org/10.4062/biomolther.2022.019
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