Molecules and Cells

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Fig. 5.

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Fig. 5. Autophagy compromise mediated by PARP1 elicits retinal degeneration in a dry AMD mouse. (A) Mice were sacrificed, and lysates of their RPE cells and retinas were analyzed with the indicated antibodies as the indicated days after SI injection. (B) Saline with 10% dimethyl sulfoxide (DMSO) or 15 mg/kg olaparib was administered via intraperitoneal injection. At 30 min after treatment, 30 mg/kg SI was injected into each mouse. At two days after injection, mice were sacrificed, and the retinas were harvested for immunoblot with the indicated antibodies. (A and B) The separated lysates between RPE cells and retinas were analyzed by immunoblotting with anti-RPE65 (marker of RPE cells). The LC3-II/-I ratio and relative p62 levels were quantified by densitometric analyses (ImageJ software). (C and D) Mice were treated with 30 mg/kg SI with or without 15 mg/kg olaparib and 1 mg/kg wortmannin by intraperitoneal injection. At five days after injection, mice were sacrificed and the retinas were harvested for histological analysis using H&E staining. Scale bars = 30 µm. (C) The black arrows indicate representative lesions in damaged RPE area. The graph shows the thickness of the outer nuclear layer (ONL) in H&E-stained samples (n = 6 eyes/group). The thickness was measured using ImageJ software. The values are presented as the mean ± SD from three independent biological replicates. Statistical analysis was performed with Student’s t-test. *P < 0.05, **P < 0.01.
Mol. Cells 2020;43:632~644
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