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1.
Figure 8

Figure 8. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

HbA1c levels in experimental animals. HbA1c levels measured at the end of the study confirmed hyperglycemic status in diabetic animals. Diabetic animals injected with fibronectin (FN)-siRNA or scrambled siRNA also displayed a similar hyperglycemic condition. *=p<0.05 versus “Normal” group mean; n=8/group.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
2.
Figure 4

Figure 4. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Effect of fibronectin (FN)-siRNA on retinal FN protein expression. A: Representative western blot image with an internal loading control β–actin showing the effect of FN-siRNA on retinal FN protein expression in diabetic rats. FN-siRNA treatment in the diabetic rats specifically reduced retinal FN protein expression compared to that in uninjected diabetic rats. B: Graphical representation of retinal FN protein expression in four groups of rats—normal, diabetic, diabetic injected with FN-siRNA, and diabetic injected with scrambled siRNA—showing that an intravitreal injection of FN-siRNA significantly reduces FN protein expression in diabetic rat retinas. Normal versus Diabetic *=p<0.05; Diabetic versus Diabetic+FN-siRNA **=p<0.05, n=8.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
3.
Figure 2

Figure 2. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Dose response study showing fibronectin (FN)–siRNA efficacy. Different doses of FN-siRNA were tested at a 1-week time point to identify an optimal dosage for effective downregulation of FN expression. The graph shows reduced FN levels in rat retinas after intravitreal injection with increasing concentrations (0.5–6.0 μM) of FN–SiRNA. A 3 μM dose was most effective at downregulating retinal FN protein expression. *=p<0.05 versus “0 μM” group mean; n=4/group.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
4.
Figure 3

Figure 3. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Time course study showing fibronectin (FN)–SiRNA efficacy. Relative levels of FN protein in rat retinas at different time points (3 days–6 weeks) after intravitreal injection of 3 μM FN–siRNA. By the 1-week time point, retinal FN protein expression was significantly reduced and remained inhibited at the 6-week time point. Scrambled siRNA showed no effect on the FN protein level. *=p<0.05 versus “0 days” group mean; n=4/group.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
5.
Figure 6

Figure 6. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Development of acellular capillaries in the diabetic retina. A-D: Representative retinal trypsin digest (RTD) images showing the effect of fibronectin (FN)-siRNA on the development of acellular capillaries (as indicated by arrows) in retinas of a (A) normal rat, (B) diabetic rat, (C) diabetic rat intravitreally injected with FN-siRNA, and (D) diabetic rat intravitreally injected with scrambled siRNA. FN-siRNA treatment in the diabetic rats specifically reduced the development of acellular capillaries compared to those of uninjected diabetic rats. E: Graphical representation of acellular capillaries in four groups of rats: normal, diabetic, diabetic injected with FN-siRNA, and diabetic injected with scrambled siRNA, showing that an intravitreal injection of FN-siRNA reduces the development of acellular capillaries in diabetic rat retinas. Normal versus Diabetic *=p<0.05; Diabetic versus Diabetic+FN-siRNA **=p<0.05, n=8.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
6.
Figure 1

Figure 1. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Localization of intravitreally injected fibronectin (FN)-SiRNA in vascular cells of a rat retina. FN-siRNA injected intravitreally in rat eyes localized on the inner limiting membrane and in retinal microvessels. The arrow points to a microvessel showing an endothelial cell nucleus (4',6-diamidino-2-phenylindole [DAPI]-positive) in panel A, and panel B shows Cyanine-3 labeled FN-siRNA localized in the same endothelial cell of the retinal microvessel. Panel C shows a superimposed image of A and B. Panel D shows a close-up view of the retinal microvessel shown in panel C. Bar: 50 μm. Inner limiting membrane (ILM); Retinal capillary (RC).

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
7.
Figure 7

Figure 7. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Retinal pericyte loss in diabetic rats. A-D: Representative retinal trypsin digest (RTD) images showing the effect of fibronectin (FN)-siRNA on the development of pericyte ghosts (as indicated by arrows) in retinas of a (A) normal rat, (B) diabetic rat, (C) diabetic rat intravitreally injected with FN–siRNA, and (D) diabetic rat intravitreally injected with scrambled siRNA. FN-siRNA treatment in the diabetic rats specifically reduced pericyte loss compared to uninjected diabetic rats. E: Graphical representation of pericyte dropouts in four groups of rats—normal, diabetic, diabetic injected with FN-siRNA, and diabetic injected with scrambled siRNA—showing that an intravitreal injection of FN-siRNA reduces the pericyte loss in diabetic rat retinas. Normal versus Diabetic *=p<0.05; Diabetic versus Diabetic+FN-siRNA **=p<0.05, n=8.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.
8.
Figure 5

Figure 5. From: A long-term siRNA strategy regulates fibronectin overexpression and improves vascular lesions in retinas of diabetic rats.

Electron microscopy (EM) analysis of capillary basement membrane (BM) thickening in rat retinas. A-D: Effect of fibronectin (FN)-siRNA on retinal capillary BM thickening in diabetic rats. Representative EM images of transverse sections of retinal capillaries of (A) normal rat, (B) diabetic rat, (C) diabetic rat intravitreally injected with FN-siRNA, and (D) diabetic rat intravitreally injected with scrambled siRNA; scale bar=1 μm. Enlarged views: E-H; scale bar=100 nm. FN-siRNA treatment in the diabetic rats specifically prevented BM thickening (arrowheads) compared to uninjected diabetic rats. (I) Graphical representation of retinal vascular BM thickness in four groups of rats: normal, diabetic, diabetic injected with FN-siRNA, and diabetic injected with scrambled siRNA, showing that an intravitreal injection of FN-siRNA reduces retinal capillary BM thickening in diabetic rats. Normal versus Diabetic *=p<0.05; Diabetic versus Diabetic+FN-siRNA **=p<0.05, n=8.

Sumon Roy, et al. Mol Vis. 2011;17:3166-3174.

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