Journal of Diabetes Research / 2016 / Article / Fig 5

Research Article

The Different Effects of Atorvastatin and Pravastatin on Cell Death and PARP Activity in Pancreatic NIT-1 Cells

Figure 5

Role of PARP1 expression in the induction of AS and PS autophagy. (a) CYTO-ID® Green Dye staining revealed significantly increased LC3-II accumulation (green) in AS-treated NIT-1 cells compared with untreated cells; decreased LC3-II accumulation was observed after treatment with the BAF-A1 inhibitor; increased LC3-II accumulation was observed after treatment with the DPQ and RAPA inhibitors. Rapamycin is included as positive control. (b) No significant difference was found in the accumulation of LC3-II between control and PS-treated or PS+BAF-A1-treated NIT-1 cells, remaining high after DPQ or RAPA pretreatment. Chloroquine is included as positive control. Magnification: ×60. Scale bar: 20 μM. (c) Quantification of the CYTO-ID Green Dye stain in NIT-1 cells per 600x fields for six fields per group. A significantly high number of LC3-II-positive cells were observed in the AS20, AS20+DPQ, and AS20+RAPA groups. (d) Cells treated with AS also showed a decreased ATP level, indicating that AS promoted ATP depletion in NIT-1 cells. DPQ significantly reduced the cellular ATP level. PS treatment for 48 h did not change the cellular ATP level and only mildly increased the ATP level after DPQ pretreatment. No significant difference was found compared with the untreated control group (, compared to the untreated group; , compared to the only AS- or PS-treated group; mean ± SD from three replicates). The value for the CON group was set at 1.
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