G9a Suppression Alleviates Corneal Neovascularization through Blocking Nox4-Mediated Oxidative Stress

Figure 3

Inhibition of G9a alleviated angiogenesis and oxidative stress through reduction of Nox4. Western blotting analysis for the expression of Nox4 and VEGF after transfection with an si-Nox4 (a-c). ROS production was measured after transfection with an si-Nox4 (d). H₂O₂ production was measured after transfection with an si-Nox4 (e). The expression of Nox4 and VEGF was evaluated through western blot after treatment with BIX (10 μM, application 2 h prior to H/R) or transfection with siRNA against G9a (f-h). ROS production was measured after treatment with BIX or transfection with siRNA against G9a (i). H₂O₂ production was measured after transfection with an si-G9a (j). HUVECs were treated with 10 μM BIX for 2 h and then infected with an adenovirus carrying the human Nox4 for 48 h, prior to exposure to the H/R process. Western blotting analysis for the expression of Nox4 and VEGF in different groups (k-m). (l,m) The production of ROS (n) and H₂O₂ (o) was measured in different groups. Data are expressed as (). versus control; ^# versus H/R; ^& versus H/R+si-NC. Experiments were repeated 3 times.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)

(l)

(m)

(n)

(o)