Fig. 4

SI induces severe lysosomal membrane permeabilization (LMP) and activation of the CLEAR network. A Representative images and quantification of immunostained LAMP-2⁺ lysosomes (gray) in ARPE-19 cells treated for 24 h, nuclei were counterstained with DAPI (blue). B Representative images and quantification of the number of cells presenting acidic (LysoTracker⁺, cyan) and/or permeabilized (Galectin-3⁺, yellow) lysosomes, nuclei were counterstained with DAPI (blue). C Diagram depicting the distinct cellular responses to lysosomal damage and LMP. D Representative images of ARPE-19 cells expressing TFEB-EGFP (green) to assess nuclear translocation, nuclei were counterstained with DAPI (blue). F Gene expression levels of genes under transcriptional control of TFEB (CLEAR network; LAMP1, CTSA, CTSD, SQSTM1) in ARPE-19 cells, evaluated by RT-qPCR. G Representative images and quantification of the number of cells with Cathepsin B activity (MagicRed, magenta), nuclei were counterstained with DAPI (blue). Scale bars, 25 μm. All data are expressed as the mean ± s.e.m. Dots represent biological replicates from three independent experiments (A) or independent experiments (B, D, F, G). P values were calculated using 1-way ANOVA with Šídák’s post-hoc test. ****P < 0.0001, ***P < 0.001, *P < 0.05, ns: not significant