C3d deposition in chronic hypoxia-induced PH in mice.

<p>(A–D) Lungs from normoxic and hypoxic C57Bl/6J mice were stained with α-C3d and α-SMA antibody and counterstained with DAPI to detect nuclei(n = 4). (E) Quantification of C3d staining in WT vs. C3−/− mice in normoxia and hypoxia. Bars represent the mean ± SD (n = 4). *<i>P</i><0.05. (F) Representative Western blot for C3d in normoxic vs. hypoxic C57Bl/6J mice. (G) Quantification of Western blots for C3d in normoxic vs. hypoxic mice. Bars represent the mean ± SD (n = 4). *<i>P</i><0.05. (H) C5a was immunoprecipitated from lung or plasma of normoxic and hypoxic C57Bl/6J mice and analyzed by Western blot (rC5a = recombinant mouse C5a).</p

Markers of Inflammation and endothelial dysfunction in WT vs. C3 −/− mice.

<p>(A) IL-6 mRNA was measured by quantitative rtPCR in RNA prepared from normoxic or hypoxic WT and C3−/− lungs. IL-6 levels were normalized to the house keeping gene β2-microglobulin. (B) ICAM-1 was quantified by ELISA in lung homogenates from normoxic or hypoxic WT or C3−/− mice. (C) ET-1 was quantified by ELISA in plasma from normoxic or hypoxic WT or C3−/− mice. Bars represent mean ± SD (n = 4) for A–C. *<i>P</i><0.05.</p

Loss of C3 prevents platelet activation caused by CH.

<p>(A) Bleeding time of WT and C3−/− mice exposed to normoxia or CH (n = 5–7). (B) Scatter plot showing platelet population in platelet rich plasma. All experiments were similarly gated to the area encircled. (C) Flow cytometry histogram demonstrating that the gated cell population is positive for the platelet marker CD41. (D–E) Representative flow cytometry histograms of platelets from (D) hypoxic WT or (E) hypoxic C3−/− mice stained with P-selectin antibody or isotype control. (F) Percent P-selectin positive platelets in PRP isolated from normoxic and hypoxic WT or C3−/− mice (n = 6). Bars represent mean ± SD. *<i>P</i><0.05; n.s. = not significant.</p

The distributions of EBC formate concentration for healthy adolescents and children with mild-to-moderate or severe asthma.

<p>Markers represent individual measurements; boxes represent the 25^th, 50^th, and 75^th percentiles; whiskers represent the mean±standard deviation.</p

Principal Component Analysis of EBC collected from all subjects.

<p>Each cluster is scaled according to the fraction of the total variance explained by that principal component. The Pearson correlation coefficients are given between all species within each cluster.</p

Loss of C3 attenuates vascular wall thickening in response to CH.

<p>(A–B) Representative photomicrographs of muscularized arterioles from (A) hypoxic WT and (B) hypoxic C3−/− mice. (C) Quantification of the % wall thickness of peripheral arterioles (<100 µm). Bars represent the mean ± SD (n = 4). *<i>P</i><0.05.</p

Characteristics of study participants¹.

1<p>Values are either frequency (percentage) or mean (range).</p>2<p>Data were logarithmically transformed prior to analysis.</p

Genetic deletion of C3 attenuates CH-induced PAH in mice.

<p>WT or C3−/− mice were exposed to CH to induce PAH. After 3 weeks (A) RVSP and (B) RVH (Fulton index), were determined. Bars represent the mean ± SD (n = 8–12). *<i>P</i><0.05.</p

C3d deposition in human PAH.

<p>(A–D) Lung sections from non-PAH or IPAH patients (n = 3) were stained with α-C3d and α-SMA antibody and counterstained with DAPI to detect nuclei. Images shown are representative (E) Quantification of C3d staining in non-PAH and IPAH patients. Bars represent the mean ± SD (n = 4). *<i>P</i><0.05.</p

Loss of C3 prevents hypoxia-induced TF upregulation.

<p>(A) Representative Western blot analysis of normoxic or hypoxic WT and C3−/− lungs for TF expression. (B) Densitometric anlysis of Western blots from (A). Bars represent the mean ± SD (n = 4 animals per group). *<i>P</i><0.05.</p