Molecules and Cells

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Fig. 3.

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Fig. 3. PARP1 downregulates autophagy via SIRT1 inhibition under H2O2 treatment in ARPE-19 cells. (A and B) ARPE-19 cells were treated with H2O2 for the indicated times (A) or for 6 h in the presence or absence of 100 µM β-nicotinamide mononucleotide (β-NMN) (B). Cont, control; Veh, vehicle. Scale bars = 10 µm. (B) ARPE-19 cells were transfected with RFP-GFP-LC3 and then seeded on poly-D-lysine-coated coverslips. Subsequently, the cells were treated with 0.5 mM H2O2 for 6 h in the presence or absence of 100 µM β-NMN. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Red and yellow arrows indicate autolysosomes and autophagosomes, respectively. (A and C) The cell lysates were immunoblotted with the indicated antibodies. The LC3-II/-I ratio and relative p62 levels were quantified by densitometric analyses (ImageJ software). (B) Quantified data are expressed as the mean ± SD from three independent biological replicates. Statistical analysis was performed by Student’s t-test. **P < 0.01, ***P < 0.001.
Mol. Cells 2020;43:632~644
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