Fig. 3. Human amylin overexpression provokes an increase in polyubiquitinated aggregates, nitrotyrosine levels, and an accumulation of depolarized mitochondria under basal state.

a INS1E (rIAPP and hIAPP) cells stimulated or not with thapsigargin (0.5 µM) were submitted to the DCFH-DA assay to detect ROS levels. b Representative immunoblot images from INS1E (WT, rIAPP, and hIAPP) cells under basal state. The histogram shows the fold increase in the ratio of nitrotyrosine/tubulin protein levels. c Quantification of the percentage of cells with depolarized mitochondria under basal state in INS1E (rIAPP and hIAPP) cells with JC-1 assay. Results are representing the mean percentage of independent experiments ± s.d. (n = 3), *P < 0.05. d Representative fluorescence microscopy images from INS1E (rIAPP and hIAPP) cells using LC3B (red) and FK2 (green) antibodies. The histogram shows the quantification of Mander’s colocalization threshold coefficient (M2) representing the mean values of independent experiments ± s.d. (n = 3), *P < 0.05