Molecules and Cells

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

Fig. 2.

Download original image
Fig. 2. PARP1 suppresses autophagy in response to H2O2 in ARPE-19 cells. (A) ARPE-19 cells were treated with 0.5 mM H2O2 for the indicated times in the presence or absence of 10 µM olaparib (Ola). Veh, vehicle. (B and C) ARPE-19 cells were transfected with scrambled siRNA (scRNA) or ATG7 targeting siRNA for 48 h. Then, the cells were treated with 0.5 mM H2O2 for 6 h in the presence or absence of 10 µM olaparib (B) for indicated times (C). (D and F) 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 for 6 h in the presence or absence of 10 µM olaparib (D) or pretreated with 100 µM rapamycin and then treated with 0.5 mM H2O2 for 6 h in the presence or absence of 10 µM olaparib (F). Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Red and yellow arrows indicate autolysosomes and autophagosomes, respectively. Cont, control. Scale bars = 10 µm. (E and F) ARPE-19 cells were pretreated with 100 µM rapamycin for 5 h and then treated with 0.5 mM H2O2 for 1 h in the presence or absence of 10 µM olaparib. (A, B, C, and E) 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). (D and F) 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, ***P < 0.001.
Mol. Cells 2020;43:632~644 https://doi.org/10.14348/molcells.2020.0078
© Mol. Cells