The corneal cauterization assay was performed, as previously described [28 (link)]. Eight-week-old female C57BL/6 mice were anesthetized with a mixture of xylazine (10 mg/kg) and ketamine (100 mg/kg). After application of a local anesthetic (Unicaïne, 0.4%) to the eye, the cornea was thermally cauterized using an ophthalmic cautery. One day after cauterization, drops of 10 µL of CXCL9(74-103) (100 µg/mL) or PBS were applied daily for 4 days. On day 5, the mice were sacrificed, and the corneas were removed. Fixation and blocking of whole-mounted corneas were performed in 70% (v/v) ethanol for 1 h and 3% (w/v) BSA in PBS for 1 h, respectively. To stain for blood vessels, the corneas were incubated with rat anti-mouse CD31 antibody (clone MEC13.3, BD Biosciences) overnight and AlexaFluor568 goat anti-rat secondary antibody (Cat No A-11077, Invitrogen) for 2 h. Corneas were flat mounted on microscope glasses overlaid with Prolong Gold antifade mounting medium and imaged using a Leica DMI6000 microscope (Leica Microsystems, Wetzlar, Germany). The cornea blood vessel area was quantified using Leica MM AF morphometric analysis software (Leica Microsystems) and expressed as the percentage of the total corneal area. Ethical approval for animal experiments was obtained from the Ethical Committee of KU Leuven (number LA1210604).
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