Rho-2A-rtTA/Tigre-TetO-hVEGFA165-IRES-EGFP

Fig.1 Detection of human VEGFA and mouse Vegfa expression in eyeball by qPCR (8wks, n=3).

Abbr. WT, Wild type; Dox, Doxycycline.

Fig.2 Detection of human VEGFA expression in serum by ELISA (8wks, n=3).

Note. The anti-human VEGFA Antibody cross-reacted with mouse Vegfa and humanized VEGFA.

Fig.3 Detection of EGFP expression in eyeball by Fundus auto-fluorescence (FAF). Bright spots were detected(red arrows), indicating the expression of EGFP. This suggests that the TetO system effectively induced the expression of the EGFP reporter gene in specific retinal cells. The expression of EGFP serves as an indicator of hVEGFA expression.

Fig.4 Fundus Fluorescein Angiography (FFA). The results showed significant vascular abnormalities in Vegfa/Rho-rtTA mice induced with Dox drinking water for two weeks, characterized by extensive fluorescein leakage(red arrows).

Fig.5 Examination of fundus and retinal morphology using optical coherence tomography (OCT). The results indicated that, compared to WT mice and non-Dox induced Vegfa/Rho-rtTA mice, Vegfa/Rho-rtTA mice exhibited structural disorganization and thicker retina, with moderate blurring and high reflectivity in the photoreceptor inner segment/outer segment (IS/OS) junction layer and part of RPE layer (red arrows) after two weeks of Dox induction through drinking water.