Acute effects of FTY720 on S1P mediated contraction.

<p>Mesenteric arteries of control animals were pre-incubated with 10μM FTY720 for 30 minutes before a dose effect curve to S1P (10nM-10μM) was constructed. S1P mediated contractions were normalized to maximal contractions to 60mM KCl. Pre-incubation with FTY720 (caused a leftward shift of the CR curve to S1P; this could be in part reversed by addition of indomethacin (10μM). n = 6 rats per group, 2 rings per rat. (*p<0.05 EC₅₀ FTY720 vs control).</p

Effect of a sub-threshold dose of S1P on myogenic constriction of endothelium-intact and -denuded mesenteric arteries of rats chronically treated with or without FTY720.

<p>Rats were treated for 6 weeks with 1mg/kg FTY720 or vehicle. Mesenteric arteries were isolated and myogenic constriction was assessed ex-vivo. (A) Pressure induced myogenic constriction of control treated animals. (B) Area under the curve of pressure induced myogenic constriction. (C) Myogenic constriction was potentiated by preincubation with 30nM S1P. (D) Area under the curve of S1P potentiated myogenic constriction. (n = 10 rats for control, n = 6 rats for FTY720, single measurement). Data are expressed as mean ± SEM. * p<0.05 vs Intact FTY720.</p

Animal body weight and hemodynamic effects of FTY720.

<p>Animal body weight and hemodynamic effects of FTY720.</p

Vascular reactivity of endothelium-intact and -denuded mesenteric arteries of rats chronically treated with or without FTY720.

<p>Rats were treated for 6 weeks with 1mg/kg FTY720 or vehicle. Mesenteric arteries were isolated and vascular constriction and relaxation assessed ex-vivo. (A) Contraction-response curve to phenylephrine (PE). (B) Relaxation-response curves to acetylcholine (Ach) in intact arteries. (C) Relaxation-response to sodium nitroprusside (SNP) in endothelium denuded arteries. (n = 4 rats per group, 2 rings per rat). Data are presented as mean±SEM.</p

Regression of lymph vessels after Ang II withdrawal, no effect on peritubular capillaries.

<p>Ang II infusion increased mRNA expression of PDPN, however this was not significant (A). Ang II-induced lymphangiogenesis was evidenced by an increase in lymph vessel formation (B), with spontaneous regression to control levels eight weeks after withdrawal of Ang II. Ang II infusion had no effects on peritublar capillaries (C). Representative photomicrographs of Podoplanin stained renal sections (D). (##p<0.01 vs. control at time of biopsy; ***p<0.001 <i>vs</i>. Ang II at time of biopsy)</p

MCP-1-induced monocyte migration is increased in the presence of immobilized heparin-albumin and glycosylated collagen XVIII.

<p><i>A:</i> MCP-1 dose dependently increased the migration of monocytes over a porous membrane. Immobilization of heparin-albumin, mimicking an artificial BM HSPG, promotes monocyte transmigration. Spontaneous migration over albumin-coated membrane in the absence of MCP-1 was set as 1 and the other values were calculated accordingly. The error bars represent SEM. <i>B:</i> Transmigration of monocytes towards MCP-1 (10 ng/ml) was increased in the presence of heparin-albumin and collagen XVIII with long GAG chains. Heparin-albumin increased the monocyte migration significantly compared to albumin coated membrane (p<0.01). N-terminal fragment of short collagen XVIII with long GAG chains promoted transmigration significantly compared to albumin and N-terminal fragment without GAG chain (both p<0.05). Relative to albumin, also the full-length short collagen XVIII promoted MCP-1-induced monocyte transmigration to some extent (not significant). Data is calculated relative to migration over albumin-coated membrane towards 10 ng/ml MCP-1. The error bars represent SEM. *: p<0.05, **: p<0.01.</p

No focal segmental glomerular sclerosis after AngII infusion.

<p>Representative photomicrographs of PAS stained renal sections (A).</p

Tubular damage is reduced in double mutant mice deficient of collagen XV and XVIII compared to the WT mice.

<p><i>A–D:</i> Paraffin-embedded sections stained with periodic acid Schiff's (PAS) reagent in sham operated WT(day 1; <i>A</i>), in sham-operated <i>Col15a1^−/−×Col18a1^−/−</i> compound mutant mice (day 1; <i>B</i>) and in WT (<i>C</i>) and <i>Col15a1^−/−×Col18a1^−/−</i> compound mutant mice (<i>D</i>) at day 5 after renal I/R. WT mice showed tubular casts, tubular widening and flattening, and loss of nuclei in tubular cells (black arrows). Such damage was not observed in double collagen mutant mice kidneys. Scale bars 50 µm. <i>E:</i> Percentage of tubular damage was determined at different time points after I/R (see methods) in the <i>Col15a1^−/−</i>, <i>Col18a1^−/−</i> and <i>Col15a1^−/−×Col18a1^−/−</i> double mutant mice compared to the WT controls (*: day 5 p<0.01 double mutant mice compare to WT). Data is presented as mean percentage ± SEM.</p

AngII infusion causes persistent glomerular desmin expression.

<p>Ang II infusion increased desmin protein levels (A). At sacrifice there was a significant increase of desmin protein in former AngII-infused rats and surprisingly in control rats as well. Compared to control rats, glomerular desmin expression of AngII-infused rats was significantly higher after stopping AngII infusion. Representative photomicrographs of desmin stained renal sections (B). (## p<0.01, ### p<0.001 <i>vs</i>. control at time of biopsy; *p<0.05 <i>vs</i>. AngII at time of biopsy; $ p<0.05 <i>vs</i>. control at time of sacrifice).</p

Effects of Ang II infusion and withdrawal on tubular damage.

<p>Havcr1 mRNA (A) and protein (B) levels were increased by Ang II infusion. After stopping Ang II infusion, recovery of proximal tubular damage was established, evidenced by control levels of Havcr1 mRNA and Kim-1 protein levels in Ang II infused rats. Representative photomicrographs of Kim-1 stained renal sections (C). (### p<0.001 vs. control at time of biopsy; ***p<0.001 vs. Ang II at time of biopsy).</p