Fig. 8.

Prolonged treatment with p120 siRNA causes further expansion of HCEC monolayers without EMT. (A) The HCEC monolayer surface area reached a plateau when treated with scRNA (d1, d2 and d3; 1.7±0.4 mm²; n = 9, P<0.05), but was continuously promoted by p120 siRNA (D1, D2 and D3) (P<0.05 on day 18 except the D3/d3 pair, and on Day 21, 24 and 27 for all three pairs; 3.7±0.7 mm²; n = 9, P<0.05) without cell enlargement in the center. (B) The HCEC density was 2241±104/mm² when the Descemet membrane was stripped from the peripheral cornea (a). It increased to 2548±93/mm² for HCEC monolayers cultured on Day 14 (b, n = 5, P<0.05). For the control treated with scRNA, the HCEC density dropped to 2083±86/mm² on Day 28 (c, n = 5, P>0.05) and 1764±96 mm² on Day 38 (d, n = 5, P<0.05), i.e. 10 days after withdrawal. By contrast, the HCEC density was maintained at 2316±79/mm² on Day 28 (i.e. two weeks of p120 siRNA3 treatment) (e, n = 5, P>0.05), and 2289±113/mm² on Day 38 (i.e. 10 days after withdrawal, f, n = 5, P>0.05). (C) Prolonged p120 siRNA treatment results in nuclear translocation of p120 and dissolution of F-actin without disturbing the junctional staining pattern of N-cadherin, ZO-1 and Na⁺/K⁺-ATPase. Ten days after withdrawal of p120 siRNA, the staining pattern of p120 and F-actin reverted to the normal pattern whereas that of the other proteins remained unchanged. Scale bars: 100 µm.