Fig. 5

gp91ds-tat treatment down-regulates the NOX2 overexpression induced by Pressure 60 mmHg-injured in the retinal vessels and the GCL. (A) Retinal ROS production was measured using DCF diacetate probe. Data are presented as the percent fluorescence intensity of the groups versus the WT-Control. (B) Representative images of DHE and CD31 co-stained retinal cryosections from WT and Nox2^−/−retinas after Pressure 60 mmHg 24 h. Scale bar, 50 μm. The white arrows point to cross-sections of retinal blood vessels. Lower panels in (B) show the enlarged views of the boxed regions (Scale bars equal 10 μm). (C) Analysis of DHE staining intensity in ECs, vessels, and GCL. Data are presented as the percent fluorescence intensity of the groups versus the control. (D) Representative images of DHE and CD31 co-stained retinal cryosections from Pressure 60 mmHg 24 h retinas with 300µM gp91ds-tat treatment. Scale bar, 50 μm. The white arrows point to cross-sections of retinal blood vessels. Lower panels in (D) show the enlarged views of the boxed regions (Scale bars equal 10 μm). (E) Analysis of DHE staining intensity in ECs, vessels, GCL, INL and ONL. Data are presented as the percent fluorescence intensity of the groups versus the control. (F) Representative images of retinal cross sections immunostained with NOX2 after Pressure 60 mmHg with 300µM gp91ds-tat treatment. The white arrows point to cross-sections of retinal blood vessels. Scale bar, 50 μm. (G) Analysis of NOX2 fluorescence intensity in the retinal vessels and GCL. Data are presented as the percent fluorescence intensity of the groups versus control-Vehicle. (H) Messenger RNA expression of Nox2 and p47^phoxin retinas after H-IOP and with 500µM gp91ds-tat treatment. Data are presented as the fold-change versus control-vehicle. (I) After H-IOP and with 500µM gp91ds-tat treatment, retinal extracts were assayed for Western blot analysis of gp91^phox and p47^phox. Representative Western blots are presented. (J) The results obtained from Western blot are expressed as the ratio to β-actin from 6–9 independent experiments. Data are shown as mean ± SEM (n = 6 in each group, one-way ANOVA with Tukey’s multiple comparisons test, *p < 0.05, **p < 0.01,***p < 0.001, ****p < 0.0001)