Fig. 4

Real-time fluorescence imaging of lysosomes and cytosolic acidosis simultaneously. For a–d, Hela cells were co-cultured with 1 (10 µM) and LysoTracker Green (0.3 µM) for 30 min, then imaged under a microscope. a Cells were excited with a 543 nm laser, and the emission was collected between 580 and 730 nm. b Cells were excited with a 488 nm laser. c A merged image combining a and b. d Pearson intensity scatter plot of ROI. For panels e–j, Hela cells were loaded with 1 for 30 min and were submitted to confocal microscope observation followed by e, f addition of 10 μM chloroquine (images at different time intervals shown; plot of fluorescence intensity over time in f. g Addition of 10 μM dexamethasone (images at different time intervals shown); plot of fluorescence intensity over time in h. i In-situ incubation in a citric acid/phosphate buffer that was adjusted to pH ~4.5 and supplemented with nigericin and KCl (140 mM) (images at different time intervals shown); plot of fluorescence intensity over time in j. Scale bar 10 μm.