Irradiating the lungs of young adult female FVB/N mice with either a single 12 Gy dose or fractionated 24 or 30 Gy doses of ¹³⁷Cs γ-rays leads to a marked decrease in the expression of aquaporin-5 in the lung.

<p>Lung tissues were collected at 14 weeks (12 Gy and 30 Gy) or at 1, 2 and 5 months (24 Gy) PI. Cell lysates were generated from the snap-frozen left lung from 4 mice and run on a 10–15% polyacrylamide mini-gel. Each lane contains the pooled lysate from 2 mice. ß-actin served as loading control. <b>A</b>: The reduction in aquaporin-5 protein at 14 weeks after a single 12 Gy or a fractionated 30 Gy dose is similar. <b>B</b>: The reduction in aquaporin-5 protein occurs as early as 1 month after a fractionated 24 Gy dose and remains reduced for the next 4 months. <b>C</b>: Quantification of the aquaporin-5 protein levels in 3B shows that the reduction in aquaporin-5 is progressive over the first 2 months postirradiation (PI) and then remains at a relatively constant low level up to 5 months PI. Densitometry was used to quantify the protein in each lane; all irradiation values in 2C were normalized to the unirradiated control values obtained on the same gel. The data in 2C are the mean ± 1 SEM.</p

Irradiating the lungs of young adult female FVB/N mice with single and fractionated doses of ¹³⁷Cs γ-rays leads to dose-dependent increases in mortality.

<p>Groups of 8–10 week-old female FVB/N mice were irradiated with single (11–13 Gy) or fractionated (18–36 Gy) doses of γ rays, and their survival recorded up to 22 weeks postirradiation.</p

Irradiating the lungs of young adult female FVB/N mice with either a single 12 Gy dose or fractionated 24 or 30 Gy doses of ¹³⁷Cs γ-rays leads to decreased levels of E-cadherin and increased levels of vimentin protein in the lung.

<p><b>A, C</b>: Although the vimentin data is variable, the decrease in E-cadherin and the increase in vimentin are similar at 14 weeks after a single 12 Gy dose or a fractionated 30 Gy dose. <b>B, D</b>: After a fractionated 24 Gy dose, there is a slight increase in E-cadherin at 1 month postirradiation (PI) followed by a progressive decrease up to 5 months PI. After a fractionated 24 Gy dose, there is a progressive increase in vimentin up to 2 month PI that remains relatively constant for the next 3 months. All methods and analyses are identical to those described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053628#pone-0053628-g003" target="_blank">Fig. 3</a>.</p

Pro-SP-c and α-SMA protein co-localize in the lungs of FVB/N mice at 14 weeks postirradiation with a single dose of 12 Gy.

<p>Lung section (5 µm) from an unirradiated control (A) and an irradiated mouse (B) were co-stained with pro-SP-c and α-SMA antibodies. In the representative photomicrographs the pro-SP-c protein is red, the α-SMA protein is green, and the nuclei are blue.</p

Irradiating the lungs of young adult female FVB/N mice with either a single 12 Gy dose or fractionated 24 or 30 Gy doses of ¹³⁷Cs γ-rays leads to increase in VCAM-1 and TGF-ß1 protein in the lung.

<p><b>A</b>: The increase in VCAM-1 and TGF-ß1 is similar at 14 weeks after a single 12 Gy dose or a fractionated 30 Gy dose. <b>B</b>: Both VCAM-1 and TGF-ß1 increase up to 2 months after a fractionated 24 Gy dose and then decrease up to 5 months postirradiation.</p

Irradiating the lungs of young adult female FVB/N mice with single and fractionated doses leads to regional fibrosis and collagen deposition and increased inflammatory cell infiltration.

<p>Female FVB/N mice were received thoracic radiation with 12 and 13 Gy single dose or fractionated dose 24 Gy. Mice were euthanized at 14 weeks (12 and 13 Gy groups) or 1, 2 and 5 months (24 Gy fractionated group) postirradiation, respectively. Lung sections were stained with H & E, Massons Trichrome and/or α-SMA. A shows representative lung images of H & E staining (control: a and d; irradiated with a single 12 Gy: b and e; 13 Gy: c and f) and Masson's trichrome staining (control: g; 12 Gy: h) for mouse treated with a single dose radiation. B shows representative lung images of H & E staining (control: a and d; 1 m postirradiation (PI) with 24 Gy fWTI: b and f; 2 m PI: c and g; 5 m PI: d and h) and α-SMA staining (control: i; 1 m PI: j; 2 m PI: k; 5 m PI: l) for mice treated with 24 Gy dose of fWTI.</p

Irradiating the lungs of young adult female FVB/N mice with either a single 12 Gy dose or fractionated 24 or 30 Gy doses of ¹³⁷Cs γ-rays leads to increases in pro-SP-c protein and pro-SP-c positive cells in the lung.

<p><b>A</b>: The increase in pro-SP-c⁺ cells is similar at 14 weeks after a single 12 Gy dose or a fractionated 30 Gy dose. <b>B</b>: The increase in pro-SP-c⁺ cells after a fractionated 24 Gy dose is progressive over the first 2 months postirradiation (PI) and then remains relatively constant up to 5 months PI. <b>C</b>: Upper gel, the increase in pro-SP-c protein at 14 weeks after a single 12 Gy or a fractionated 30 Gy dose is similar. Lower gel, pro-SP-c protein is increase by 1 month PI and remains relatively constant up to 5 months PI. For 4A and 4B, lung tissues were collected and prepared for histochemical analysis either at 14 weeks (12 Gy and 30 Gy) or 1, 2 and 5 months (24 Gy) PI. Sections (5 µm) of lung tissue were stained with pro-SP-c antibodies, and the pro-SP-c⁺ cells were analyzed using morphometric method. Data are the mean ± 1 SEM; n = 3; ^*<i>p</i><0.05 vs. sham control. The gels in 4C were prepared and analyzed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053628#pone-0053628-g003" target="_blank">Fig. 3</a>.</p