CFB−/− mice on a C57BL/6 background were generously provided by V. Michael Holers (University of Colorado Health Science Center, Denver, CO). C57BL/6 mice were generated from breeding pairs (Harlan Laboratories, Indianapolis, IN). Animals were housed under a 12-hour light/12-hour dark cycle with access to food and water ad libitum.
For CNV lesions, 3-month-old mice were anesthetized (20 mg/kg xylazine, 80 mg/kg ketamine), and pupils were dilated (2.5% phenylephrine HCl and 1% atropine sulfate). Argon laser photocoagulation (532 nm, 100 μm spot size, 0.1 second duration, 100 mW) was used to generate four laser spots in each eye surrounding the optic nerve with the use of a handheld coverslip as a contact lens. Bubble formation at the laser spot indicated the rupture of Bruch membrane.17 (link)
For tail-vein injections, the vein was vasodilated by heat, a 25-gauge needle was inserted, and a 50 μL volume was injected (250 μg CR2-fH in PBS, molar equivalent of CR2 in PBS [113 μg], or PBS only). Dosing and treatment schedules are outlined in Results. All experiments were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the University Animal Care and Use Committee.
For CNV lesions, 3-month-old mice were anesthetized (20 mg/kg xylazine, 80 mg/kg ketamine), and pupils were dilated (2.5% phenylephrine HCl and 1% atropine sulfate). Argon laser photocoagulation (532 nm, 100 μm spot size, 0.1 second duration, 100 mW) was used to generate four laser spots in each eye surrounding the optic nerve with the use of a handheld coverslip as a contact lens. Bubble formation at the laser spot indicated the rupture of Bruch membrane.17 (link)
For tail-vein injections, the vein was vasodilated by heat, a 25-gauge needle was inserted, and a 50 μL volume was injected (250 μg CR2-fH in PBS, molar equivalent of CR2 in PBS [113 μg], or PBS only). Dosing and treatment schedules are outlined in Results. All experiments were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the University Animal Care and Use Committee.