Molecules and Cells

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Fig. 4.

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Fig. 4. The PARP1-SIRT1 axis participates in mitochondrial dysfunction upon H2O2 treatment in ARPE-19 cells. (A and E) ARPE-19 cells were treated with 0.5 mM H2O2 in the presence or absence of 100 µM β-nicotinamide mononucleotide (β-NMN) for 3 h. Then, the mitochondrial membrane potential of the cells was measured using a Muse analyzer (A) or intracellular ATP using a VICTOR plate reader (E). The cells in the left quadrant are depolarized cells. (B) The graph was obtained from a quantitative analysis of depolarized cells. (C) ARPE-19 cells were transfected with DsRed-Mito to observe mitochondrial morphology (red) for 48 h and seeded on poly-D-lysine-coated coverslips. Subsequently, the cells were treated with 0.5 mM H2O2 in the presence or absence of 100 µM β-NMN for 1 h. The nuclei were visualized by 4′,6-diamidino-2-phenylindole (DAPI) staining. Cont, control. Scale bars = 10 µm. (D) The graph shows the numbers of fragmented mitochondria per cells (n ≥ 10). (B, D, and E) 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
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