Organ elimination half-life of exogenous mouse sRAGE administered by three different routes.

a<p>Not determined due to ambiguity, interruption, or lack of convergence of fit.</p

Mouse and human sRAGE extinction coefficients.

a<p>Mouse membrane RAGE has a published sequence length of 402 (NP_031451.2).</p>b<p>Human membrane RAGE has a published sequence length of 404 (NP_001127.1).</p>c<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088259#pone.0088259-Pace1" target="_blank">[50]</a>.</p

Kinetics of binding of extracellular matrix proteins in solution to solid-phase sRAGE.

a<p>Not determined due to undetectable specific binding.</p

Clearance of intratracheally-administered sRAGE or MSA from mouse lung is rapid and non-proteolytic.

<p>(A) Exogenous mouse sRAGE or MSA persistence in mouse lung as a function of time. (B) SDS-PAGE separation and gel autoradiography of lung homogenates from mice administered i.t. mouse sRAGE or MSA and sacrificed at the indicated time points thereafter.</p

RAGE binds with high affinity to collagen I, collagen IV, and laminin, but not to fibronectin.

<p>Soluble RAGE affinity for collagen I, collagen IV, laminin, and fibronectin was assessed by biolayer interferometry. Soluble RAGE was conjugated to an amine-reactive sensor and dipped into solutions containing collagen I (A), collagen IV (B), laminin (E), or fibronectin (F) for 1800 s, at which point dissociation was initiated by washing in PBS. Binding was measured as a deflection in the wavelength of light passing through the sensor; affinities were calculated by fitting the curves with analysis software. Reciprocal binding was studied with collagen I (C) and collagen IV (D) conjugated to the sensor and sRAGE in solution. Laminin and fibronectin could not be conjugated to the sensor.</p

sRAGE and MSA used in biodistribution and clearance studies are very pure.

<p>(A) SDS-PAGE separation and Coomassie Brilliant Blue staining of ∼2.5 µg of MSA or mouse sRAGE preparations used for radiolabeling. (B) SDS-PAGE separation and gel autoradiography of ¹²⁵I-labeled MSA and mouse sRAGE.</p

Organ biodistribution of intravenously-administered sRAGE or MSA in mice.

<p>Organ biodistribution of mouse sRAGE or MSA (A) 1 hour, (B) 2 hours, (C) 4 hours, and (D) 12 hours after intravenous injection of ∼1 µCi of tracer, corresponding to ∼0.6–1.4 µg of radiolabeled protein. Asterisks indicate statistically significant differences.</p

Organ biodistribution of intraperitoneally-administered sRAGE or MSA in mice.

<p>Organ biodistribution of mouse sRAGE or MSA (A) 1 hour, (B) 2 hours, (C) 4 hours, and (D) 12 hours after intraperitoneal injection of ∼1 µCi of tracer, corresponding to ∼0.6–1.4 µg of radiolabeled protein. Asterisks indicate statistically significant differences.</p

Organ biodistribution of intratracheally-administered sRAGE or MSA in mice.

<p>Organ biodistribution of mouse sRAGE or MSA (A) 1 hour, (B) 2 hours, (C) 4 hours, and (D) 12 hours after intratracheal instillation of ∼1 µCi of tracer, corresponding to ∼0.6–1.4 µg of radiolabeled protein. Asterisks indicate statistically significant differences.</p

Clearance of intratracheally-administered sRAGE from mouse lung occurs at the alveolar-capillary interface.

<p>Representative light micrographs/autoradiographs recorded at 20× magnification of sections of lung from mice given mouse sRAGE (or MSA, shown for comparison) i.t. and sacrificed at the indicated time points thereafter. Lung from untreated mice serves as a control for background radiation.</p