cDNA nucleotide sequences and expression of the genes encoding the cytoplasmic ribosomal proteins S18 and S27 from the green alga Chlamydomonas reinhardtii

Path length (cm) measured during the probe trial in the Morris water maze (MWM). Mice were let in the MWM for a 1-min free swimming test. Total distances traveled were quantified and showed no difference between all tested groups (15 F− A+/+, 14 F+ A+/+, and 11 F+ A+/−)

Additional file 2: of Activation of phagocytic activity in astrocytes by reduced expression of the inflammasome component ASC and its implication in a mouse model of Alzheimer disease

Path length (cm) measured during the probe trial in the Morris water maze (MWM). Mice were let in the MWM for a 1-min free swimming test. Total distances traveled were quantified and showed no difference between all tested groups (15 F− A+/+, 14 F+ A+/+, and 11 F+ A+/−)

Lung accumulation of Th 17 cells but not T regs is limited in the absence of MyD88 signalling.

<p>Lymphocyte numbers in the bronchoalveolar lavage (BAL) fluid (<b>A</b>) and CD4+ T lymphocyte numbers in lung suspensions (<b>B</b>) obtained from wild-type (WT) and MyD88-knockout (MyD88-KO) mice 60 days after silica (SiO₂) or saline solution (NaCl) treatment. Expression of Foxp3 (<b>C</b> and <b>D</b>), IL-17A (<b>E</b>) and RORγ (<b>F</b>) analyzed by RTqPCR in lungs (<b>C</b>) and in lung CD4+ T lymphocytes (<b>D–F</b>) obtained from WT and MyD88-KO mice 60 days after silica or saline treatment. Expression of Foxp3 and IL-17A was normalized on that of 18S RNA. Bars represent means ± SEM (n = 3–5). These results were treated statistically by a t-test. ns indicates no statistically significant difference and * = p<0.05 and ** = p<0.01 indicates statistically significant difference between values measured in silica-treated WT mice and silica-treated KO mice.</p

Robust fibrotic response in the absence of innate immune receptors or ligands.

<p>Hydroxyproline (OH-proline) content assessed in lung homogenates of mice 60 days after saline (NaCl) or silica (SiO₂) instillation in wild-type (WT), IL-1R- (<b>A</b>), ASC- (<b>B</b>), NALP3- (<b>C</b>), IL-1β- (<b>D</b>), TRIF- (<b>E</b>), TLR2/4- (<b>F</b>), TLR3- (<b>G</b>), γδTCR- (<b>H</b>), IL-23p19- (<b>I</b>) knockout (KO) mice. Bars represent means ± SEM (n = 3–5). These results were treated statistically by a t- test. ns indicates no statistically significant difference and ** = p<0.01 indicates statistically significant difference between values measured in silica-treated WT and silica-treated KO mice.</p

Lung inflammation but not collagen accumulation is reduced in MyD88-KO mice treated with silica particles.

<p>Neutrophil (<b>A</b>) and macrophage (<b>B</b>) numbers in the bronchoalveolar lavage (BAL) fluid of wild-type (WT) and MyD88-knockout (MyD88-KO) mice treated with silica (SiO₂, 2.5 mg/mouse) or saline solution (NaCl) and sacrificed at day 60. Quantification of IL-1β (<b>C</b>) by ELISA in the BAL fluid of WT and MyD88-KO mice treated with SiO₂ or NaCl at day 60. Lung fibrosis assessed by OH-proline contents (<b>D</b>) in WT and MyD88-KO mice after silica or saline instillation (d60). Bars represent means ± SEM (n = 4–6). These results were treated statistically by a t- test. ns indicates no statistically significant difference and ** = p<0.01 indicate statistically significant difference between values measured in silica-treated WT and silica-treated KO mice.</p

MyD88-KO mice treated with silica particles developed limited granuloma and neutrophil infiltration but marked fibrosis and lymphocyte accumulation.

<p>5-µm sections of paraffin-embedded lung tissue of wild-type (WT) and MyD88- knockout (MyD88-KO) mice treated with silica (SiO₂, day 60) were stained with hematoxylin and eosin (panels <b>A–B</b> and <b>I–J</b>; neutrophils are indicated by black arrows and lymphocyte aggregates by an asterisk), with Masson's trichrome (panels <b>C–F</b>) or with silver (panels <b>G–H</b>). Sections are representative of 3–4 mice examined. Magnification 50X (panels A and B), 100X (panels <b>C–H</b>) and 400X (panels I–J). Histological quantification of fibrotic nodule, diffuse fibrosis and lymphoid aggregate numbers (<b>K</b>). These results were treated statistically by a t- test. * = p<0.05, ** = p<0.01 indicate statistically significant difference between values measured in silica-treated WT and silica-treated KO mice.</p

Pulmonary accumulation of T regs and Th17 lymphocytes is associated with the development of silica-induced lung fibrosis.

<p>Number of CD4+ (<b>A</b>) and CD8+ (<b>B</b>) analyzed by flow cytometry in lung cell suspensions obtained from wild-type mice after silica (SiO₂, d3 to d60) or saline (d0) instillation. Expression of Foxp3 (<b>C</b>) and IL-17A (<b>D</b>) analyzed by RTqPCR in the lungs obtained from wild-type mice after silica (d3 to d60) or saline (d0) treatment. Expression of Foxp3 and IL-17A was normalized on that of 18S RNA. Bars represent means ± SEM (n = 3–5). These results were treated statistically by a Student Neuman-Keul's test. * = p<0.05, ** = p<0.01, *** = p<0.001 indicate statistically significant difference between values measured in saline-treated mice and silica-treated mice.</p

Lung expression of TGF-β1, IL-10 and PDGF-B is not reduced in MyD88-KO mice treated with silica particles.

<p>Lung expression of TGF-β1 (<b>A</b>), IL-10 (<b>B</b>) and PDGF-BB (<b>C</b>) analyzed by RTqPCR in tissue or by ELISA in BAL of wild-type (WT) and MyD88-knockout (MyD88-KO) mice 60 days after silica (SiO₂) or saline (NaCl) treatment. Expression of PDGF B (<b>D</b>) analyzed by RTqPCR in lung CD4+ T lymphocytes purified from NaCl- or SiO₂-treated WT and MyD88-KO mice at day 60. Expression of TGF-β1, IL-10 and PDGF-B was normalized on that of 18S RNA. Bars represent means ± SEM (n = 3–5). These results were treated statistically by a t- test and no statistically significant difference (ns) between values measured in silica-treated WT mice and silica-treated KO mice was noted.</p

Exaggerated proliferation and myofibroblast differentiation of CF fibroblasts.

<p>Cell proliferation and myofibroblast differentiation in cultured lung (A–D) and skin (E–H) fibroblasts at the second passage purified from CF mice homozygous for the F508del mutation and from wild-type (WT) mice. A,e) Uptake of³H-thymidine (1 µCi/well) assessed in cultured cells seeded at 30×10³ cells/well, in the absence of any added growth factor to culture media or in the presence of 1 to 100 ng/ml human rPDGF-BB for 1 h. After 48 h, adherent cells were trypsinized before ³H-thymidine counting. Data expressed as counts per minute (cpm). b,f) Cell growth analysis assessed by daily counting, in a Neubauer chamber, of trypsinized cells cultured in the absence of any added growth factor to culture media. c,g) α-SMA mRNA expression, using 18S RNA as a reference, assessed in the absence of any added growth factor to culture media or in the presence of 1 to 100 ng/ml human rTGF-β1 for 24 h. d) α-SMA mRNA expression assessed in the presence of 0.1 to 50 µM vardenafil (Vard) for 6 h. h) Micrographs of fibroblast cultures under stimulation with 10 ng/ml human rTGF-β1. Arrows identify formation of cellular aggregates. Bars: 100 µm. Values are means ± SEM of 3 multi(96)well cultures per group from a representative experiment selected from at least 3 experiments with similar results. *: <i>P</i><0.05; **: <i>P</i><0.01; *** <i>P</i><0.001 for comparison of mean values.</p

Vardenafil prevents overresponsive proinflammatory status in CF fibroblasts.

<p>a,b–g) mRNA and c) protein expression of proinflammatory cytokines in response to 0.1 mg/ml LPS in lung (a,b,e–g) and skin (c,d) cultured fibroblasts at the second passage purified from CF mice homozygous for the F508del mutation and from wild-type (WT) mice. At the mRNA level, markers were assessed 3 h after LPS stimulation. At the protein level, CCL-2 (c) was assessed 24 h after LPS stimulation. Vardenafil (Vard; 0.1 µM) was used for TNF-α (e), IL-1β (f) and IL-6 (g) mRNA expression studies. 18S RNA used as a reference gene. Values are means ± SEM of 3 multi(96)well cultures per group from a representative experiment selected from at least 3 experiments with similar results. *: <i>P</i><0.05; **: <i>P</i><0.01; *** <i>P</i><0.001 for comparison of mean values.</p