Figure 4.

AAV5-Id3 gene therapy regulates profibrotic genes and regulates ECM organization

(A–D) qRT-PCR data reflect that AAV5-Id3 gene therapy attenuates the mRNA expression of profibrotic genes (α-smooth muscle actin (α-SMA], fibronectin [FN], collagen I [Col-I], and collagen III [Col-III], respectively) in corneal tissues compared with AAV5-naked vector control group corneal tissues on day 28. (E–G) Immunofluorescence staining of fibrosis marker α-SMA. Images of corneal tissue sections show that AAV5-Id3 gene therapy governs the fibrotic response at the protein level compared with AAV5-naked vector control group corneal tissues on day 28. (H) Quantification graph shows that AAV5-Id3 gene therapy significantly reduces the α-SMA level compared with AAV5-naked vector control group corneal tissues. (I–K) Masson’s trichrome staining images of corneal tissue sections show that AAV5-Id3 gene therapy regulates collagen formation and maintains collagen-ECM organization compared with the AAV5-naked vector control group corneal tissues, as evident from a higher level of blue color intensity on day 28. (L) Quantification graph shows that AAV5-Id3 gene therapy significantly reduces the collagen level compared with AAV5-naked vector control group corneal tissues. Tissues were collected on day 28. There were six samples in each group, and error bars represent ±SEM. ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. Scale bars, 100 μm.