Effect of 5-LO products on the total cortical TGF-β expression increased during kidney injury.

<p>Mice were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107549#s2" target="_blank">Materials and Methods</a> section (<i>n</i> = 6 per group). (A) Representative immunohistochemical staining for total TGF-β in cortical areas of WT mice and 5-LO-deficient mice treated with saline or BSA (bars  = 40 µm). Arrows indicates markedly positive staining. (B) Quantitative analyses were expressed as means ± SE. Statistically significant in relation to *WT + SAL (<i>p</i><0.05).</p

Nestin expression is increased in subcapsular and corticomedullary glomeruli of mice subjected to kidney injury.

<p>Mice were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107549#pone-0107549-g001" target="_blank">Fig. 1</a> (<i>n</i> = 6 per group). (A) Representative immunohistochemical staining for nestin in the subcapsular glomerulus and (B) corticomedullary glomerulus of WT and 5-LO-deficient mice treated with saline or BSA (bars  = 20 µm). Quantitative analyses (C,D) were expressed as means ± SE. Statistically significant in relation to *WT+SAL (<i>p</i><0.05), #WT+BSA (<i>p</i><0.05), ⁺5-LO^–/–+SAL (<i>p</i><0.05).</p

Macrophage infiltration is attenuated in 5-LO-deficient mice subjected to kidney injury.

<p>Mice were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107549#pone-0107549-g001" target="_blank">Fig. 1</a> (<i>n</i> = 6 per group). Representative immunohistochemical staining for F4/80 in (A) cortical and (B) medullary areas of WT mice and 5-LO-deficient mice treated with saline or BSA (bars  = 40 µm). Quantitative analyses (C,D) were expressed as means ± SE. Statistically significant in relation to *WT+SAL (<i>p</i><0.05), #WT+BSA (<i>p</i><0.05), ⁺5-LO^–/–+SAL (<i>p</i><0.05).</p

LTs modulate albumin uptake and kinase activity.

<p>LLC-PK1 cells were grown on 6-well plates, kept overnight in medium depleted of serum in the presence of 10⁻⁷ M LTD₄ or 10⁻⁷ M LTB₄. After treatment, both (A) albumin uptake and (B) PKC activity were measured as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107549#s2" target="_blank">Materials and Methods</a> section. The cells were preincubated with 10⁻⁸ M calphostin C (Calph C) when indicated. (C) The effect of LTs on PKB activity measured by Ser473 phosphorylation. PKC (D) and PKB (E) activities were measured in WT and 5-LO-deficient mice treated with saline or BSA. The results are expressed as means ± SE. Statistically significant in relation to *control or WT+SAL (<i>p</i><0.05), #WT+BSA (<i>p</i><0.05).</p

Proteinuria and urinary tubular enzymes are attenuated in 5-LO-deficient mice subjected to kidney injury.

<p>The animals were given i.p. injections of saline (SAL; 0.9%) or 10 g/kg BSA for 7 days as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107549#s2" target="_blank">Materials and Methods</a> section. (A) The proteinuria was measured on days 0, 2, 4, and 6. (B) Urine samples were resolved on SDS-PAGE gels and protein analysis was based on the intensity of Coomassie Blue staining. (C) LDH was measured in urine samples as an index of cell damage. WT+SAL (<i>n</i> = 8), WT+BSA (<i>n</i> = 8), 5-LO^–/–+SAL (<i>n</i> = 8), 5-LO^–/–+BSA (<i>n</i> = 8). The results are expressed as means ± SE. Statistically significant in relation to *WT+SAL (<i>p</i><0.05), #WT+BSA (<i>p</i><0.05), ⁺5-LO^–/–+SAL (<i>p</i><0.05), and ^&5-LO^–/–+BSA (<i>p</i><0.05).</p

Basic features of BSA-induced tubulointerstitial injury model.

<p>(A) Representative Periodic-acid Schiff staining in renal cortex of WT+SAL and WT+BSA groups. Arrows, indicates epithelial cell vacuolization; *, indicates tubular shedding; &, indicates disorganization of brush-border. (B) Quantification of tubulointerstitial space area. Results were expressed as means ± SE. Statistically significant in relation to *WT + SAL (<i>p</i><0.05).</p

Renal function parameters.

<p>Statistically significant in relation to *WT+SAL (day 7; <i>p</i><0.05), ⁺5-LO^–/–+SAL (day 7; <i>p</i><0.05).</p><p>Renal function parameters.</p

Glomerular morphology in WT and in 5-LO-deficient mice after 7 days of treatment with saline or BSA.

<p>Mice were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107549#pone-0107549-g001" target="_blank">Fig. 1</a> (<i>n</i> = 6 per group). Kidney sections were stained with periodic acid–Schiff. (A) Representative photomicrographs of the subcapsular glomeruli (bar  = 40 µm) and (B) corticomedullary glomeruli (bar  = 40 µm). Quantitative analysis of (C) the glomerular tuft surface of the subcapsular glomerulus, (D) the glomerular tuft surface of the corticomedullary glomerulus, (E) the mesangial surface of the subcapsular glomerulus, and (F) the mesangial surface of the corticomedullary glomerulus. The results are expressed as means ± SE. Statistically significant in relation to *WT+SAL (<i>p</i><0.05), #WT+BSA (<i>p</i><0.05), ⁺5-LO^–/–+SAL (<i>p</i><0.05).</p