Research Paper Volume 13, Issue 24 pp 25670—25693

Figure 4. LC3B particle analysis in polarized ARPE-19 cells upon MTK and ZTK treatment. Representative IF images of polarized ARPE-19 cells in (A) the absence and (B) presence of lysosomal inhibitors E64d and pepstatin A for 3 h and the (C, D) corresponding LC3B particle count and size evaluation by ImageJ using the Yen thresholding method. Relative E64d/pepstatin A-induced LC3B particle (E) count accumulation, (F) size increase and (G) particle-associated LC3B levels/cell (particle count x size) of control, MTK- and ZTK-treated polarized ARPE-19 cells grouped in low and high E64d/pepstatin A-induced particle accumulation of control samples. Values are represented in box and whisker plot format (min to max); independent experiments: n = 4–5. The significance of differences in relative E64d/pepstatin A-induced LC3B particle count, size and count x size upon MTK and ZTK treatment was calculated by a repeated measures two-way ANOVA (main factors: low and high E64d/pepstatin A-induced particle accumulation and MTK/ZTK treatment (matched)) followed by a Dunnett multiple comparison test. ^**p < 0.01, ^*p < 0.05 compared to the control, ⁺⁺p < 0.01 compared to the low E64d/pepstatin A-induced particle accumulation sample.