FIG 5.

AZT incubation leads to lysosomal hyperactivation. C2C12 cells were incubated in the absence or presence of AZT (6, 30, 150 µM) or 3TC (8, 40, 200 µM) for up to 8 days. (A) Flow cytometry analysis of total lysosomal abundance using LysoTracker Red (LTR) (left) with representative histograms (right). (B) Flow cytometry analysis of lysosomal acidity using LysoSensor Green (LSG) (left) with representative histograms (right). (C and D) Flow cytometry analysis of cathepsin L (D) and B (C) enzymatic activity using Magic Red cathepsin L and B assay kits (left) with representative histograms (right). The results of all experiments are presented as the mean ± SD and are representative of those from at least three independent experiments with four replicates. ***, P < 0.001.