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1.
Figure 6.

Figure 6. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Nitrotyrosine staining increased after bleomycin treatment in WT mice, but not in mice overexpressing EC-SOD. (A) Relative densitometry of nitrotyrosine signal in lung homogenates from WT and EC-SOD TG mice. *P < 0.05 according to one-way ANOVA versus WT PBS-treated mice (n = 6 per group). (B) Representative image of slot blot for lung nitrotyrosine content in WT and EC-SOD TG mice with PBS treatment, and 21 and 35 days after bleomycin treatment.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
2.
Figure 7.

Figure 7. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

eNOS expression decreased in WT mice at the latest time point, but increased in mice overexpressing EC-SOD. (A) Relative densitometry for endothelial nitric oxide synthase (eNOS) protein expression relative to β-actin was determined by immunoblot, using a rabbit polyclonal eNOS antibody (1:2,000). *P < 0.05 according to one-way ANOVA versus WT PBS-treated mice. #P < 0.05 according to one-way ANOVA versus WT 35D mice (n = 4 per group). (B) Representative image of eNOS expression in each group, taken from a single blot.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
3.
Figure 8.

Figure 8. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Bleomycin-induced pulmonary hypertension and mortality was attenuated in mice overexpressing EC-SOD. (A) Right ventricular systolic pressures (RVSP; mm HG) in WT mice and EC-SOD TG mice 21 and 35 days after intratracheal treatment with a single dose (0.1 unit) of bleomycin, compared with control mice treated with PBS. *P < 0.05 versus PBS-treated mice according to one-way ANOVA. #P < 0.05 versus WT 35D mice (n = 8–12). (B) A Kaplan-Meier survival curve shows weekly survival over a 35-day period in the WT mice (solid line) and EC-SOD TG mice (dashed line). *P < 0.05 according to log rank test.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
4.
Figure 5.

Figure 5. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Redox-sensitive transcription factor early growth response (Egr-1) and transforming growth factor (TGF-β) gene expression increased 7 days after bleomycin treatment in WT lungs, but not in lungs of mice overexpressing EC-SOD. mRNA expression for TGF-β (A) and Egr-1 (B) was measured by RT–quantitative PCR in lung homogenates of WT and EC-SOD TG mice 7 days after bleomycin treatment, compared with sham-treated control mice (n = 6 in each group). Data are expressed as copies of gene per copy of hypoxanthine-guaninie phosphoribosly transferase (HPRT), standardized to sham WT mice. *P < 0.05 compared with PBS-treated WT mice. #P < 0.05 versus WT 7D, according to one-way ANOVA.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
5.
Figure 1.

Figure 1. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Pulmonary artery medial wall thickening developed by 35 days after intratracheal bleomycin treatment in wild-type (WT) mice but not in mice overexpressing extracellular superoxide dismutase in the lung (EC-SOD TG). (A) In lung sections stained with α–smooth muscle actin (α-SMA) to define the medial wall, medial wall thickening (mwt) was measured in pulmonary arteries (PAs) < 50 μm in control PBS-treated mice, and 21 and 35 days (21D and 35D, respectively) after bleomycin treatment. *P < 0.05 versus intratracheal control mice by one-way ANOVA. We counted 7–10 vessels per lung, with n = 4 mice per group. (B) Representative images of PAs stained with α- SMA and counterstained with hematoxylin. Arrows indicate pulmonary artery.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
6.
Figure 4.

Figure 4. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Pulmonary artery intimal thickening developed by 35 days after intratracheal bleomycin treatment in WT mice, but not in mice overexpressing EC-SOD. (A) Fifty vessels were counted in each lung after co-staining with α-SMA and Von Willebrand factor VIII, to identify the medial and intimal layers. Intimal thickening was defined as an intimal layer occupying more than 50% of the diameter of the intima + lumen. Data are shown for WT and EC-SOD TG mice 35 days after bleomycin treatment. *P < 0.05 versus 35D bleomycin-treated WT mice according to one way ANOVA. n = 3–4 per group. (B) Representative images of lung sections were co-stained with α-SMA (red) and Von Willebrand factor VIII (green), and counterstained with DAPI to identify the nuclei (blue). Arrows indicate pulmonary artery.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
7.
Figure 3.

Figure 3. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Proliferation was detected within the pulmonary artery wall 21 days after intratracheal bleomycin treatment in WT mice but not in mice overexpressing EC-SOD. (A) Sections were dual-stained with α-SMA and proliferating cell nuclear antigen (PCNA), and counterstained with 4′6-diamidino-2-pheylindole (DAPI). The percentages of proliferating cells within the pulmonary artery, identified by PCNA staining within the nuclei of small pulmonary arteries (< 100 μm), were determined in WT and EC-SOD TG mice at 21 days after bleomycin treatment. *P < 0.05 versus PBS-treated mice. #P < 0.05 versus 21D bleomycin-treated mice according to one-way ANOVA (n = 3–4 mice per group). (B) Representative images of pulmonary arteries co-stained with α-SMA (red) and PCNA (green). Left: α-SMA and DAPI (blue). Right: Same vessel with PCNA and DAPI. Arrows indicate a positive nucleus; scale bar is 50 microns.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.
8.
Figure 2.

Figure 2. From: Lung Extracellular Superoxide Dismutase Overexpression Lessens Bleomycin-Induced Pulmonary Hypertension and Vascular Remodeling.

Pulmonary artery adventitial collagen deposition was increased 21 days after intratracheal bleomycin treatment in WT mice, and was attenuated in mice overexpressing EC-SOD. Computer analysis determined percent collagen deposition in whole lung (A) and within the pulmonary artery (B), as shown for WT and EC-SOD TG mice 21 days and 35 days after bleomycin treatment. *P < 0.05 according to one-way ANOVA versus PBS-treated mice. #P < 0.05 versus WT mice at comparable time points (n = 3–4 mice per group). (C) Representative images of collagen deposition by trichrome staining around pulmonary artery. Arrows indicate pulmonary artery. (D) mRNA expression for COL1A1 was measured by RT–quantitative PCR in lung homogenates of WT and EC-SOD TG mice 7 days after bleomycin treatment, compared with sham-treated controls (n = 6 in each group). Data are expressed as copies of gene per copy of HPRT standardized to the sham WT. *P < 0.05 compared with PBS-treated WT mice. #P < 0.05 versus WT 7D, according to one-way ANOVA.

Zachary Van Rheen, et al. Am J Respir Cell Mol Biol. 2011 April;44(4):500-508.

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