Journal: Clinical and Experimental Allergy

Article Title: A protective role for periostin and TGF-β in IgE-mediated allergy and airway hyperresponsiveness

doi: 10.1111/j.1365-2222.2011.03840.x

Figure Lengend Snippet: Periostin protein increases in the lung following airway challenge with A spergillus antigen. (a) Periostin protein expression in the lungs of wild-type (Wt) and periostin deficient ( P n −/− ) mice was determined by immunoblotting for periostin protein in lung extracts from saline or A spergillus (Asp) antigen-challenged mice (F3). (b) Immunohistochemistry for periostin protein expression was performed on lung sections from Wt or P n −/− mice treated with saline or Asp antigen (F3). Magnification: 40×.

Article Snippet: CD4 + CD25 − cells were stimulated for 4 days with plate-bound anti-CD3 (2 μg/mL; 145-2C11, NA/LE, BD Bioscience), soluble anti-CD28 (2 μg/mL; BD Bioscience) and recombinant human IL-2 (20 U/mL, NCI Preclinical Repository, Frederick, MD, USA) and were cultured at 10 6 cells/mL with recombinant human TGF-β1 (10 ng/mL, Humanzyme, Chicago, IL, USA) and recombinant human periostin (40 ng/mL, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, Saline, Immunohistochemistry

Journal: Clinical and Experimental Allergy

Article Title: A protective role for periostin and TGF-β in IgE-mediated allergy and airway hyperresponsiveness

doi: 10.1111/j.1365-2222.2011.03840.x

Figure Lengend Snippet: Periostin deficient mice have increased airway hyperreactivity and higher systemic IgE responses following airway challenge with A spergillus antigen. (a) Airway reactivity to intravenously administered acetylcholine is similar in unchallenged periostin deficient ( P n −/− ) mice (F6) and wild-type (Wt) mice, but acetylcholine reactivity following A spergillus (Asp) challenge is more severe in P n −/− mice (F6). * P < 0.05, *** P < 0.001 for Wt Asp vs. P n −/− Asp. ### P < 0.001 Wt saline vs. Wt Asp. (b) Serum IgE levels are similar in P n −/− mice (F6) and wild-type mice, but IgE levels following Asp challenge are more severe in P n −/− mice (F6). * P < 0.05. (c) Total cell numbers in broncho-alveolar lavage (BAL) are similar in P n −/− mice (F6) and wild-type mice, and increased markedly following Asp challenge with increases in macrophages, eosinophils and lymphocytes; these increases occurred similarly in P n −/− (F6) and wild-type mice. Results represent mean ± SEM for 5–7 mice per group (F6).

Article Snippet: CD4 + CD25 − cells were stimulated for 4 days with plate-bound anti-CD3 (2 μg/mL; 145-2C11, NA/LE, BD Bioscience), soluble anti-CD28 (2 μg/mL; BD Bioscience) and recombinant human IL-2 (20 U/mL, NCI Preclinical Repository, Frederick, MD, USA) and were cultured at 10 6 cells/mL with recombinant human TGF-β1 (10 ng/mL, Humanzyme, Chicago, IL, USA) and recombinant human periostin (40 ng/mL, R&D Systems, Minneapolis, MN, USA).

Techniques: Saline

Journal: Clinical and Experimental Allergy

Article Title: A protective role for periostin and TGF-β in IgE-mediated allergy and airway hyperresponsiveness

doi: 10.1111/j.1365-2222.2011.03840.x

Figure Lengend Snippet: Periostin deficient mice and wild-type controls have a similar increase in GFP + IL -4 producing cells and GFP + CD4 + cells within the lung following airway challenge with A spergillus antigen. Schematic scheme showing a representative flow cytometric analysis for GFP + cells (a) or GFP + CD4 + cells (c) performed on whole lung single-cell suspensions from periostin deficient mice ( P n −/− ) in C57BL/6 4get background (F6) and wild-type (Wt) littermate controls. There is a similar increase in GFP-producing cells (b) and GFP + CD4 + cells (d) in the lungs of P n −/− mice and Wt littermate controls following A spergillus (Asp) antigen challenge. Results represent mean ± SEM for 3–5 mice (F6) per group. ** P < 0.01.

Article Snippet: CD4 + CD25 − cells were stimulated for 4 days with plate-bound anti-CD3 (2 μg/mL; 145-2C11, NA/LE, BD Bioscience), soluble anti-CD28 (2 μg/mL; BD Bioscience) and recombinant human IL-2 (20 U/mL, NCI Preclinical Repository, Frederick, MD, USA) and were cultured at 10 6 cells/mL with recombinant human TGF-β1 (10 ng/mL, Humanzyme, Chicago, IL, USA) and recombinant human periostin (40 ng/mL, R&D Systems, Minneapolis, MN, USA).

Techniques:

Journal: Clinical and Experimental Allergy

Article Title: A protective role for periostin and TGF-β in IgE-mediated allergy and airway hyperresponsiveness

doi: 10.1111/j.1365-2222.2011.03840.x

Figure Lengend Snippet: Periostin deficient mice have less TGF-β1 and Foxp3 gene transcript and TGF-β1 protein in the lung following A spergillus antigen challenge. TGF-β1 and Foxp3 mRNA (F6) and TGF-β1 protein (F3) in lungs from saline and A spergillus (Asp) antigen-challenged, wild-type (Wt) and P n −/− mice. (a) TGF-β1 transcripts increase in Wt mice following allergen challenge but do not in the P n −/− mice. (b) TGF-β1 protein per 20 μg of total protein increases in Wt mice following allergen challenge but does not in the P n −/− mice. (c) Foxp3 transcripts are decreased in P n −/− mice compared with wild-type (Wt) controls following allergen challenge. Results represent mean ± SEM for 3–5 mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: CD4 + CD25 − cells were stimulated for 4 days with plate-bound anti-CD3 (2 μg/mL; 145-2C11, NA/LE, BD Bioscience), soluble anti-CD28 (2 μg/mL; BD Bioscience) and recombinant human IL-2 (20 U/mL, NCI Preclinical Repository, Frederick, MD, USA) and were cultured at 10 6 cells/mL with recombinant human TGF-β1 (10 ng/mL, Humanzyme, Chicago, IL, USA) and recombinant human periostin (40 ng/mL, R&D Systems, Minneapolis, MN, USA).

Techniques: Saline

Journal: Clinical and Experimental Allergy

Article Title: A protective role for periostin and TGF-β in IgE-mediated allergy and airway hyperresponsiveness

doi: 10.1111/j.1365-2222.2011.03840.x

Figure Lengend Snippet: Epithelial cell-derived periostin induces Foxp3 expression in T cells in a TGF-β1 dependent manner. Recombinant human TGF-β1 (10 ng/mL) but not recombinant human periostin induces CD25 expression (a) and Foxp3 gene expression (c) in activated CD4 + CD25 − mouse T cells. Co-culture of activated CD4 + CD25 − mouse T cells with Beas2B cells over-expressing periostin (B2BPN) but not a control vector (B2BCTL) induces CD25 expression (b) and Foxp3 gene expression (d). The induction of Foxp3 gene expression in activated CD4 + CD25 − mouse T cells co-cultured with B2BPN is blocked by treatment with the TGF-β1 receptor kinase inhibitor SB-431542 (SB) and the pan-TGF-β blocking antibody (BA) (e). Results represent mean ± SEM, N = 3. * P < 0.05, ** P < 0.01.

Article Snippet: CD4 + CD25 − cells were stimulated for 4 days with plate-bound anti-CD3 (2 μg/mL; 145-2C11, NA/LE, BD Bioscience), soluble anti-CD28 (2 μg/mL; BD Bioscience) and recombinant human IL-2 (20 U/mL, NCI Preclinical Repository, Frederick, MD, USA) and were cultured at 10 6 cells/mL with recombinant human TGF-β1 (10 ng/mL, Humanzyme, Chicago, IL, USA) and recombinant human periostin (40 ng/mL, R&D Systems, Minneapolis, MN, USA).

Techniques: Derivative Assay, Expressing, Recombinant, Gene Expression, Co-Culture Assay, Control, Plasmid Preparation, Cell Culture, Blocking Assay

Journal: Clinical and Experimental Allergy

Article Title: A protective role for periostin and TGF-β in IgE-mediated allergy and airway hyperresponsiveness

doi: 10.1111/j.1365-2222.2011.03840.x

Figure Lengend Snippet: Periostin deficient mice develop increased epithelial mucin stores and peribronchial collagen deposition in response to A spergillus antigen. (a) Representative periodic acid-Schiff (PAS)-stained sections from saline treated wild-type (Wt) or periostin deficient ( P n −/− ) mice demonstrate minimal PAS-positive staining. A spergillus (Asp) antigen treated Wt and P n −/− mice show magenta staining within epithelial cells, representing mucin stores. Magnification: 20×. (b) Epithelial mucin stores, represented as the volume of PAS-stained regions referenced to the area of epithelial basal lamina, increase markedly after Asp challenge in both P n −/− mice and wild-type mice (F3). Results represent mean ± SEM for 5–6 mice (F3) per group. (c) Representative Sirius red-stained sections from saline treated Wt or P n −/− mice demonstrate minimal collagen staining. Asp treated Wt and periostin P n −/− mice show increased peribronchial Sirius red staining representing airway fibrosis. Magnification: 20×. (d) Peribronchial fibrosis of small-medium sized airways (100–300 μm diameter), represented as the volume of Sirius red staining per basal lamina, increase after Asp challenge in both P n −/− mice and Wt mice (F3). Results represent mean ± SEM for 3–4 mice (F3) per group. Results represent mean ± SEM for (F3) per group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: CD4 + CD25 − cells were stimulated for 4 days with plate-bound anti-CD3 (2 μg/mL; 145-2C11, NA/LE, BD Bioscience), soluble anti-CD28 (2 μg/mL; BD Bioscience) and recombinant human IL-2 (20 U/mL, NCI Preclinical Repository, Frederick, MD, USA) and were cultured at 10 6 cells/mL with recombinant human TGF-β1 (10 ng/mL, Humanzyme, Chicago, IL, USA) and recombinant human periostin (40 ng/mL, R&D Systems, Minneapolis, MN, USA).

Techniques: Staining, Saline

Journal: bioRxiv

Article Title: Cardiac fibroblasts regulate cardiomyocyte hypertrophy through dynamic regulation of type I collagen

doi: 10.1101/2022.05.25.493406

Figure Lengend Snippet: (A) Whole ventricle mRNA microarray analysis of Col1a2 -/- mouse hearts compared to Col1a2 +/- at 2 months of age, n=3 per genotype. (B) Mass spectrometry analysis of ECM protein changes in Col1a2 -/- mouse hearts compared to Col1a2 +/- hearts at 3 months of age, n=4 per genotype. (C) Representative immunofluorescence images and (D) Western blot analysis of periostin from hearts of Col1a2 +/- and Col1a2 -/- mice at 3 months of age. Scale bar: 25 µm. (E) Flow cytometric gate strategy and (F) analysis of cardiac fibroblasts (MEFSK4 + /CD31 - /CD45 - ) from dissociated hearts of Col1a2 +/- and Col1a2 -/- mice at 3 months of age. (G) Representative immunofluorescence images of platelet-derived growth factor receptor (PDGFR)-α (purple) in Col1a2 +/- and Col1a2 -/- mice at 3 months of age. Wheat germ agglutinin (WGA) staining is green and shows outlines of cardiomyocytes. Scale bar: 100 µm. Relative mRNA expression of Col1a2 (H), Postn (I), Col3a1 (J) and Col5a1 (K) in sorted cardiac fibroblasts (MEFSK4 + /CD31 - /CD45 - ) from Col1a2 +/- and Col1a2 -/- mice at 9 months of age. Student t -test for panels (F), (H), (I), (J) and (K).

Article Snippet: Antibodies against the following proteins were used: periostin (Novus Biologicals NBP1-30042; 1:300 dilution for IF, 1:1000 for Western blot); collagen I (Abcam ab21286; 1:100 for IF); PDGFRα from (R&D Systems AF1062; 1:1000 for IF); collagen 1a2 (Santa Cruz sc-393573; 1:500 for Western blot) Anti-CD31 was from BioLegend (102423; 1:100 for flow cytometry); anti-CD45 was from BD Biosciences (563890; 1:100 for flow cytometry); anti-MEFSK4 was from Miltenyi Biotec (130-120-802; used 1:30 for flow cytometry).

Techniques: Microarray, Mass Spectrometry, Immunofluorescence, Western Blot, Derivative Assay, Staining, Expressing

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Upregulation of Periostin and Reactive Stroma Is Associated with Primary Chemoresistance and Predicts Clinical Outcomes in Epithelial Ovarian Cancer.

doi: 10.1158/1078-0432.CCR-14-3111

Figure Lengend Snippet: Figure 3. POSTN correlates desmoplasia phenotype in vivo and promotes chemoresistance in EOC cells in vitro. A, increased desmoplasia is correlated with POSTN expression and primary chemoresistance. Representative high magnification images of H&E staining for tumor specimens (top) and POSTN ISH images (bottom) are shown. Desmoplasia scores were defined as follows: 0, no desmoplasia; 1, few scattered desmoplastic foci abutting cancer cells; 2, several desmoplastic foci abutting cancer cells or moderate confluent (wider) desmoplasia, but not present throughout the section; 3, desmoplastic reaction throughout section, associated with most cancer cells. DS, desmoplastic stroma; NS, normal stroma; TC, tumor cells. Arrows, examples of tumor cells. Dotted line encircles a region containing tumor cells. Size bars, 100 mm. B, summary of desmoplasia scores in 21 Plat-S primary, 18 Plat-R primary, and 21 Plat-R recurrent tumor specimens. C, POSTN promotes chemoresistance in chemosensitive ES-2 ovarian cells in vitro. Ninety-six- well plates were coated with recombinant protein FN1 or POSTN or left uncoated before cells were plated into each well. Then, 10 mmol/L carboplatin or 10 nmol/L Taxol was added to each well on the next day. CellTiter-Glo reagents were added at 72 hours after compound treatment to measure cell viability. The viability in coated wells was then compared with the one in uncoated wells to calculate percentage growth benefit.

Article Snippet: Ninety-six-well plates were first coated with recombinant full-length FN1 (Cat# F2006; Sigma-Aldrich), POSTN (Cat# 3548-F2; R&D Systems) or left uncoated at 37 C for 2 hours or 4 C for 16 hours.

Techniques: In Vivo, In Vitro, Expressing, Staining, Recombinant