Fig. 4: MNU induces lysosomal alterations that are not reversed by DFP.
From: Deferiprone protects against photoreceptor degeneration by inhibiting parthanatos
a Representative images of ARPE-19 cells treated with different doses of MNU (500 and 1000 μg/ml) and labeled with LysoTracker Red (red, to label acidic lysosomes) and DAPI (blue). Insets show a cell treated with vehicle or 1000 μg/ml MNU (right). b Quantification by fluorescence microscopy of acidic lysosomes in cells treated with 100, 500, or 1000 μg/ml MNU. c Representative images of ARPE-19 cells treated with different doses of MNU (500 µg/ml, 1000 µg/ml) and labeled with LysoSensor Yellow/Blue DND-160, which allows distinction of basic lysosomes (blue; top) from acidic lysosomes (yellow; bottom). Fluorescence microscopy quantification of basic lysosomes (d), acid lysosomes (e), and basification ratio (f) in cells treated with 100, 500 or 1000 µg/ml MNU. g Representative images of ARPE-19 cells treated with 500 or 1000 µg/ml MNU or LLOMe (positive control) and transfected with a fluorescent Galectin-3 (green) reporter. Cells were also stained with DAPI (blue). h % of cells expressing the reporter (Gal3 (+)), detected by fluorescence microscopy, relative to the total number of cells. Dots represent different cell experiments. All data are expressed as the mean ± SEM. *p < 0.05, **p < 0.01; ***p < 0.001; ***p < 0.001. Statistical analyses were performed using a two-tailed Student´s t-test. Scale bars: 25 µm.
