Figure 4

GA101 induces lysosomal cell death (a) and senescence (b) in MALC. (a) Left panel: Detection of lysosomal membrane permeabilization after acridine orange (AO) staining of MALC treated or not with 10 μg/ml antibodies for 20 days. Box shows overlay of fold changes compared with UT MALC for one representative experiment. Histograms represent the mean fold change ±s.d. of five independent experiments. *P<0.05. Analysis and representation were performed with Cytobank. Insert: 2D RL culture cells treated with 50 μM chloroquine or 50 nM bafilomycin for 3 or 24 h were used as positive controls for lysosomal cell death. Right panel: MALC were treated (10 μg/ml antibodies) or not for 20 days, and cathepsin D expression was determined by western blot analysis. Results are representative of three independent experiments. β-Actin expression was used as a control of protein expression. (b) Detection of cleavage of C₁₂FDG by SA-βGal was measured by an increase in green fluorescence in treated and UT MALC after 20 days of culture. 2D RL culture cells treated with 2.2 μM cisplatin or 0.9 μM etoposide for 3 days were used as a positive control of senescence. Left: Overlays of fold changes comparing treated to UT cells in one representative experiment. Right: Histograms represent the mean fold change ±s.d. of five independent experiments *P<0.05. Analysis and representation were performed with Cytobank.