(A) Adult wild-type mice were bilaterally injected with either hSEPT9/eGFP-LV or control/eGFP-LV. Mice injected with hSEPT9/eGFP-LV exhibited consistent and significant IOP elevation compared with those injected with control/eGFP-LV. Viral vector injections were repeated after 2 weeks. n = 10–12 eyes. *P < 0.05 based on the nonparametric Wilcoxon’s rank-sum test. Data are presented as mean ± SEM; mouse experiments were repeated twice. (B) Expression of recombinant protein in the AH outflow pathway was confirmed in mice injected with the viral vectors through eGFP fluorescence detection. (C) Light microscopy–based histological examination of the AH outflow pathway tissues revealed no notable differences (except the SC lumen area) between the hSEPT9/eGFP-expressing and control (eGFP-expressing) mice. (D–F) Immunofluorescent staining of the AH outflow pathway tissues (especially the TM) showed increased levels of fibronectin, collagen IV, α-SMA, p-MLC, and F-actin in mice expressing hSEPT9/eGFP compared with control specimens. Representative images are shown (n = 4). (F) Quantification of immunofluorescence in the trabecular pathway (TM and inner wall of SC) revealed a significant increase in F-actin, p-MLC, α-SMA, collagen IV α1, and fibronectin in hSEPT9/eGFP-expressing mice compared with control mice. Tissue sections were stained with Hoechst (nuclei staining in blue). Representative ROI tracing used for the TM and inner wall of SC fluorescence quantification is shown in E (see the F-actin panel for reference). n = 4. *P < 0.05, **P < 0.01 based on Student’s t test. Scale bars: 50 μm (B and C) and 20 μm (D and E). TM, trabecular meshwork; SC, Schlemm’s canal; CB, ciliary body; LV, lentiviral vector; CLV, control lentiviral vector; S9LV, SEPT9-expressing lentiviral vector.