Fig 4.

A) ARPE-19 cells were transiently co-transfected with Ub-RFP and LC3—GFP for 24 hours. Appropriate treatments of cysteamine (250 μM), fisetin (40 μM), and 10% CSE were incubated with the cells for 12 hours before image capture. CSE treatment showed an observable increase in yellow bodies, indicating LC3-GFP and Ub-RFP co-localization, which is ameliorated with cysteamine or fisetin treatment. B) The number of yellow bodies per image were counted and expressed as a percentage of control. CSE treatment induces a statistically significant (p < 0.05) increase in the number of Ub-RFP and LC3-GFP co-localized bodies, while treatment with cysteamine (250 μM) and fisetin (40 μM) both showed a significant reduction (p value < 0.05) in the number of co-localized Ub-RFP and LC3-GFP aggresome bodies. From this data, we can conclude that CSE induces aggresome body formation in ARPE-19 cells through autophagy-impairment and treatment with cysteamine and fisetin reduces the formation of aggresome bodies. Data is presented as ± SEM. * p < 0.05 ** p < 0.01 with n = 4 samples per group and two images taken per group.