*4.1. Animal Experiments*

Male Sprague–Dawley rats (Charles River) weighing 180 g to 250 g were housed in a facility with 1:1 light/dark cycle. The animals were acclimated for at least 7 days and had free access to food and water. The well-established model of puromycin aminonucleoside nephropathy (PAN) was achieved by injecting puromycin aminoglycoside dissolved in 0.9% saline (125 mg/kg body weight i.p.). Rats injected with 0.9% saline served as controls. Eight days later, renal afferent lymphatic vessels were harvested for pressure myography. In a separate subset of PAN and control rats, renal lymph was collected using a glass pipette. The animal protocol was approved by Vanderbilt University Medical Center Institutional Animal Care and Use Committee in accordance with National Institutes of Health guidelines.
