FIG. 2.

Mitochondrial and nuclear DNA damage in FECD. All three cell lines were treated with 25 μM or 50 μM MN for 2 h or pretreated with NAC in the presence of 100 μM MN for 2 h and assessed for DNA damage using long-amplicon qPCR. Control (0) included DMEM for 2 h. Data represented in (A) DNA amplification of the small mitochondrial amplicon (250 bp), (B) DNA amplification and lesion frequency of mitochondrial target amplicon (8.9 kb), and (C) DNA amplification and lesion frequency of β-globin (nuclear) target amplicon (13.5 kb) (n = 3). Relative amplification was calculated comparing treated samples with untreated HCEnC-21T cells. Panel (D) depicts DNA amplification and lesion frequency in pooled normal (n = 22 for mtDNA and n = 15 for nDNA) and FECD (n = 31 for mtDNA and n = 11 for nDNA) specimens. Data are expressed as the mean ± SE. Two-way ANOVA was applied for statistical significance comparing treated samples with untreated control (0). (**p < 0.01, ***p < 0.001, p ≠ < 0.01 FECDi/FECD vs. normal cells [HCEnC-21T, HCECi] or normal donor corneal endothelium). mtDNA, mitochondrial DNA; NAC, N-acetyl-cysteine.