Relationship between serum PTH concentration and the capacity of kidney to reabsorb phosphate normalized for the glomerular filtration rate (TmP/GFR) in 112 control subjects (diamonds).

<p>The patient with the mutation in NHERF1 gene is represented by a circle.</p

Effect of mutant NHERF1 on the cell surface expression of NPT2a assessed by biotinylation experiments.

<p>Hela cells were transfected with GFP-tagged-NPT2a alone or together with a plasmid containing the cDNA of the wild type NHERF1 or the E68A mutant. NPT2a proteins labelled with sulfo-NHS-SS-biotin were revealed by using anti-GFP antibodies. Labeled proteins were isolated with avidin-agarose beads according to manufacturer's manual (Pierce). Proteins eluted from the beads and total cell lysate were loaded on a SDS gel. Western blot was probed with anti-GFP antibodies. Na+ K+ APTase was used as a loading control and detected with specific antibodies. The higher panel shows a western blot representative of four experiments. Lower panel: Statistical analyses of the quantification of the western blot (n = 4). Results are presented as mean ±sd. Quantification of the surface expression of NPT2a: Anova p = 0.038. Post Hoc test: Tukey-Kramer multiple comparisons test (#: p<0.05; * p<0.01).</p

Biochemical data of the patient.

<p>Biochemical data of the patient.</p

Measurement of phosphate transport in cells expressing the wild type or the E68A mutant NHERF1 together with the sodium phosphate cotransporter NPT2a.

<p>Panel A: Phosphate-induced current recorded in Xenopus oocytes injected with water or with either cRNA of NPT2a alone or in association with the cRNA of the wild type (WT) NHERF1 or the E68A mutant. Results are means ± SD, n = 9. Overall comparison was performed with the use of the Kruskall-Wallis test (P<0.0001) then the group expressing NPT2a alone was compared with other groups with the use of the Mann-Whitney test (p<0.01). Panel B: Sodium-dependent phosphate uptake was measured in Hela cells transfected with empty plasmids or with the plasmid containing the cDNA of NPT2a alone or together with the cDNA of the wild type or the NHERF1 mutant. Results are means ± SD, n = 6. Overall comparison was performed with the use of the Kruskall-Wallis test (P<0.0001) then the group expressing NPT2a alone was compared with the other groups with the use of the Mann-Whitney test (* p<0.01 vs NPT2a+WT NHERF1, # p <0.001 vs NPT2a alone).</p

Co-immunoprecipitation of NHERF1 with NPT2a in Hela cells.

<p>Hela cells were transfected with a plasmid containing the cDNA of NPT2a tagged with GFP at its N- extremity. Cells were also transfected with a plasmid containing the wild type cDNA of NHERF1 or one mutant (E68A or R153Q or E225K) tagged with Flag. Upper panel: Flag-tagged-NHERF1 protein was immunoprecipitated by using an anti-Flag antibody and the immunoprecipitates were probed with anti-GFP antibodies to detect GFP-tagged-NPT2a on Western blot. Middle panel: The total expression of GFP-tagged-NPT2a was analyzed in the cell lysates by anti-GFP Western Blot. Lower panel: Wild type and mutant Flag-NHERF1 was immunoprecipitated and probed by using anti-Flag antibodies to compare Flag-NHERF1 immunoprecipitation in the different conditions.</p

Additional file 1: of Promoter hypermethylation of HS3ST2, SEPTIN9 and SLIT2 combined with FGFR3 mutations as a sensitive/specific urinary assay for diagnosis and surveillance in patients with low or high-risk non-muscle-invasive bladder cancer

Table S1. Oligonucleotides for QM-MSP. (DOC 37 kb

The <i>MUC5B</i> Variant Is Associated with Idiopathic Pulmonary Fibrosis but Not with Systemic Sclerosis Interstitial Lung Disease in the European Caucasian Population

<div><p>A polymorphism on the <i>MUC5B</i> promoter (rs35705950) has been associated with idiopathic pulmonary fibrosis (IPF) but not with systemic sclerosis (SSc) with interstitial lung disease (ILD). We genotyped the <i>MUC5B</i> promoter in the first 142 patients of the French national prospective cohort of IPF, in 981 French patients with SSc (346 ILD), 598 Italian patients with SSc (207 ILD), 1383 French controls and 494 Italian controls. A meta-analysis was performed including all American data available. The T risk allele was present in 41.9% of the IPF patients, 10.8% of the controls (P = 2×10^–44), OR 6.3 [4.6–8.7] for heterozygous patients and OR 21.7 [10.4–45.3] for homozygous patients. Prevalence of the T allele was not modified according to age, gender, smoking in IPF patients. However, none of the black patients with IPF presented the T allele. The prevalence of the T risk allele was similar between French (10%) and Italian (12%) cohorts of SSc whatever the presence of an ILD (11.1% and 13.5%, respectively). Meta-analysis confirmed the similarity between French, Italian and American cohorts of IPF or SSc-ILD. This study confirms 1) an association between the T allele risk and IPF, 2) an absence of association with SSc-ILD, suggesting different pathophysiology.</p></div

Characteristics of the populations investigated for meta-analysis.

*<p>some data may be missing but data expressed represent always more than 2/3 of the populations. na not available.</p

Meta-analysis of the <i>MUC5B</i> rs35705950 T allele risk in the three American cohorts and the French cohort of idiopathic pulmonary fibrosis (IPF).

<p>Forrest plots of the meta-analysis of the association of the <i>MUC5B</i> rs35705950 T polymorphism and IPF. Bars represent the 95% interval.</p

Characteristics of the SSc patients in the 2 investigated European populations.

<p>Anti-Topo I: anti-topoisomerase I antibodies; ACA: anti-centromere antibodies; ILD interstitial lung disease. Data are expressed as % or mean ± SD. SSc systemic sclerosis. ILD Interstitial lung disease.</p