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. 2014 Jun 26;5(14):5547–5558. doi: 10.18632/oncotarget.2139

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Copyright: © 2014 Vasiliauskas et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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A). BrdU immunostaining was performed on prostates from 30-week HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice. Prostates from 30-week old HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice did not exhibit significant differences in BrdU incorporation. Data are expressed as the mean ± SE. Three separate areas were counted from three independent specimens per group, and representative images are shown. Arrows depict a few of the positive cells stained for BrdU. Scale bar=20um. B). Detection of TUNEL-positive cells in prostate tissue of HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice at 30-weeks of age. Prostates from 30-week old HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice stained significantly different for TUNEL. Data are expressed as the mean ± SE. *P<0.05. Three separate areas were counted from four independent specimens per group, and representative images are shown. Arrows depict a few of the positive cells stained for TUNEL. Scale bar=20um. C). Cleaved Caspase-3 immunostaining was performed on prostates from 30-week HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice. Prostates from 30-week old HGFL-/- TRAMP+ mice had significantly higher Cleaved Caspase-3 staining. Data are expressed as the percent positive prostate epithelial cells ± SE. *P<0.05. Four separate areas were counted from three independent specimens per group, and representative images are shown. Arrows depict a few of the positive cells stained for Cleaved Caspase-3. Scale bar=20um. D). Western blot analyses of whole prostate tumor lysates from HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice for downstream Ron signaling targets. E). HGFL-/- TRAMP+ mice have decreased vascularization as determined by CD31 staining compared to HGFL+/+ TRAMP+ mice. Bar graphs depict mean vessel density per area with n=3 mice per each group and 3 random fields per mouse. Representative images are shown. Data are expressed as the mean ± SE. *P<0.05. Scale bar=20um. F). F4/80 immunostaining was performed on prostates from 30-week HGFL+/+ TRAMP+ and HGFL-/- TRAMP+ mice. F4/80-positive cells infiltrated prostate tumors from HGFL-/- TRAMP+ mice whereas F4/80-positive cells localized on the tumor periphery in prostate tumors from HGFL+/+ TRAMP+ mice.