Figure 5.

Knockdown (KD) of Neat1 expression in the mouse corneal endothelium and in human corneal endothelial cells

(A) Experimental timeline including intracameral injection of rAAV and FECD model construction. rAAV 4wk: UVA irradiation 4 weeks post-injection of virus. (B) Representative confocal images of whole mount of corneal endothelium detecting ZO-1 treated with control shRNA (NC) and Neat1 shRNA (Neat1 KD) in normal and FECD mouse. Scale bars, 50 μm. (C) ZO-1 immunostaining-based analysis for cell density, hexagonality, and coefficient of variation (n = 3). (D) Representative OCT images of mouse corneas at 3 months post-UVA. (E) OCT image-based CCT analysis from pre-UVA to 3 months post-UVA. n = 3 for each group. (F) Cell viability of HCEC upon NEAT1 KD with or without H₂O₂ treatment. (G) Mitochondrial abundance by comparative quantitative real-time-PCR upon KD of NEAT1. (H) Cellular ROS levels in NEAT1 KD HCEC upon H₂O₂ treatment. Quantitative data are shown as means ± SEs, n = 3. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; and ∗∗∗∗p < 0.0001 (2-tailed t test).