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. 2019 Dec 6;2019:5637075. doi: 10.1155/2019/5637075

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Copyright © 2019 Audrey Voisin et al.

This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Analysis of hiPSC-RPE lysosomal activity during oxidative stress induced by Fe-NTA treatment. (a) Flow cytometry analysis and (b) quantification of global lysosomal activity in both hiPSC-RPE Control (N = 5) and AMD (N = 6) cells (one analysis per cell line). Statistical analysis: two-way ANOVA and post hoc Bonferroni. (c) Western blot analysis of pro-Cat D and mature Cat D and (d) western blot quantification in both hiPSC-RPE Control cells (N = 6) and AMD cells (N = 6) (one analysis per cell line). Statistical analysis: two-way ANOVA and post hoc Fisher LSD. Comparison of each dose between hiPSC-RPE Control and AMD cells (^∗). ^∗p < 0.05. (e) Analysis of hiPSC-RPE Control cells (N = 6) and AMD cells (N = 6) cell Cat B activity (four analyses per cell line). Statistical analysis: two-way ANOVA and post hoc Bonferroni. Comparison of each dose between hiPSC-RPE Control and AMD cells (^∗). ^∗p < 0.05 and ^∗∗p < 0.01. (f) Comparison of the ratio of the dimer or oligomer to the monomer of acridine orange in hiPSC-RPE Control (N = 6) and AMD (N = 5) cells (triplicate per cell line). ^∗Comparison of each dose between hiPSC-RPE Control and AMD cells for each dose. ^$Comparison of each dose between basal condition for hiPSC-RPE Control and AMD cells. Statistical analysis: two-way ANOVA and post hoc Bonferroni. ^∗^,$p < 0.05, ^∗∗^,$$p < 0.01, and ^∗∗∗^,$$$p < 0.0001. (g) Lysosomal membrane stability was measured by AO staining under a fluorescence microscopy.