Gyc 500

Induction of laser-induced choroidal neovascularization (LCNV) was performed in six adult C57BL/6 mice, three of each sex, following published protocols (20 (link), 21 , 34 (link)). Briefly, following anesthetization and pupillary dilation, four laser-induced choroidal lesions were created in each eye by rupturing the Bruch’s membrane with a solid-state laser photocoagulator, GYC-500 (Green photocoagulation 532 nm laser) mounted on a slit-lamp (Nidek, Fremont, CA, USA). Lesions were created using the following laser parameters: 100 μm spot size, 0.1 s duration, and 0.1 Watts. Rupture of the Bruch’s membrane was confirmed using OCT imaging (Phoenix Research Laboratories, Pleasanton, CA, USA) of the mouse LCNV as shown in Supplementary Figure 6. Four days later, the LCNV mice were injected intraperitoneally with 10 mg/kg InflammaProbe-1 or an equimolar dose of Oregon Green 488 cadaverine, 5-isomer (5.46 mg/kg) in 100 μL PBS with 10% DMSO. Brightfield and fluorescent fundus images were acquired 6 h post-injection using the Micron IV retinal imaging system (Phoenix Research Laboratories, Pleasanton, CA, USA). Annotations were added to both images using PowerPoint (Microsoft, Redmond, WA, USA). The images were representative of 12 eyes.

C57BL/6J mice underwent anesthesia using a 4:1 mixture of Zoletil (Virbac, Carros Cedex, France) and xylazine (Bayer Healthcare, Leverkusen, Germany) with added topical 0.5% proparacaine (Alcaine; Alcon, Geneva, Switzerland) for topical anesthesia. Pupils were dilated by Tropherine eye drops (0.5% tropicamide and 0.5% phenylephrine hydrochloric acid; Hanmi Pharm, Seoul, Republic of Korea). A slit-lamp delivery system (SL-1800; NIDEK, Tokyo, Japan) with a green laser photocoagulator (GYC-500; NIDEK) (532 nm laser, 50 μm spot size, 0.1 s duration, 200 mW) was used to generate three laser spots in each eye using 12 mm-diameter microscope cover glasses (Paul Marienfeld GmbH, Lauda-Königshofen, Germany) as contact lenses while protecting the optic nerve with a lubricant (hypromellose; SAMIL, Seoul, Republic of Korea). Successful Bruch’s membrane disruption was indicated by gaseous bubbles. After photocoagulation, intravitreal injections of GST, mCXCR3-WT (2 µg/µL), mCXCR3-S2 (2 µg/µL), and aflibercept (EYLEA, 2 µg/µL; Bayer Healthcare) were administrated. Only spots with accompanying bubbles were included in the study.