Molecules and Cells

Indexed in /covered by CAS, KoreaScience & DOI/Crossref:eISSN 0219-1032   pISSN 1016-8478

Fig. 1.

Download original image
Fig. 1. H2O2 induces PARP1 activation and compromises autophagy in ARPE-19 cells. (A and B) ARPE-19 cells were treated with 0.1 mM or 0.5 mM H2O2 for the indicated times. (A) Cell death was analyzed by flow cytometry using propidium iodide staining. (B) The cell lysates were immunoblotted with the indicated antibodies. (C) ARPE-19 cells were treated with 0.5 mM H2O2 in the presence or absence of the autophagy inhibitors 3-methyladenine (3-MA, 10 µM) and bafilomycin A1 (BafA1, 100 nM). The cell lysates were immunoblotted with the indicated antibodies. Cont, control. (B and C) LC3-II/-I ratio was quantified by densitometric analyses. (D) ARPE-19 cells were transfected with RFP-GFP-LC3 and seeded on poly-D-lysine-coated coverslips. Next, the cells were treated with 0.5 mM H2O2 or autophagy modulators (inducer: rapamycin, 100 nM, or inhibitor: BafA1, 100 nM) for 6 h. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Red and yellow arrows indicate autolysosomes and autophagosomes, respectively. Scale bars = 10 µm. (A and D) Quantified data are expressed as the mean ± SD from three independent biological replicates. Statistical analysis was performed by Student’s t-test. *P < 0.05, **P < 0.01.
Mol. Cells 2020;43:632~644 https://doi.org/10.14348/molcells.2020.0078
© Mol. Cells