Journal: Acta biochimica et biophysica Sinica

Article Title: Characterization of a micro-roughened TiO2/ZrO2 coating: mechanical properties and HBMSC responses in vitro.

doi: 10.1093/abbs/gmu040

Figure Lengend Snippet: Figure 4. Real-time PCR measurement of mRNA levels of HBMSCs cultured on TiO2/ZrO2 and TiO2 coatings in osteogenic medium and standard medium (A) Expression level of Runx2 in cells cultured in osteogenic medium was higher than that in cells cultured in standard medium at all time points. Cells cultured on TiO2/ZrO2 surfaces showed a higher Runx2 mRNA expression level than those on TiO2 coating in osteogenic medium at day 10 (*P , 0.05). (B) Expression level of Osterix in cells cultured in osteogenic medium was higher than that in cells cultured in standard medium at all time points. Cells cultured on TiO2/ZrO2 surfaces showed a higher Osterix mRNA expression level than those on TiO2 coating in osteogenic medium at days 7 and 10 (*P , 0.05). (C) Expression level of ALP in cells cultured in osteogenic medium was higher than that in cells cultured in standard medium at most time points (*P , 0.05). (D) Expression level of OPN in cells cultured in osteogenic medium was higher than that in cells cultured in standard medium at all time course (*P , 0.05).

Article Snippet: After co-incubation with primary rabbit polyclonal anti-human osteopontin antibodies (1 : 100; Proteintech Group, Chicago, USA) and primary mouse monoclonal anti-human Runx2 antibodies (1 : 100; ProSci, Poway, USA) overnight at 48C, all the samples were then washed with PBS and co-incubated with Cy3-labeled goat anti-rabbit IgG (1:1000; Beyotime, Beijing, China) and fluorescein (FITC)-labeled goat anti-mouse IgG (1:1000; Beyotime) for 1 h at RT.

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Expressing

Journal: Acta biochimica et biophysica Sinica

Article Title: Characterization of a micro-roughened TiO2/ZrO2 coating: mechanical properties and HBMSC responses in vitro.

doi: 10.1093/abbs/gmu040

Figure Lengend Snippet: Figure 5. Immunofluorescence for in vitro osteogenic differentiation of HBMSCs Immunofluorescence labeling of Runx2 (FITC, Green), OPN (Cy3, Red) and nucleus (DAPI, Blue) of HBMSCs on TiO2/ZrO2 (A–E) and TiO2 coatings (F–J) cultured in osteogenic/standard medium for 7 days. HBMSCs cultured on TiO2/ZrO2 coating (A: DAPI image, B: Cy3 image, C: FITC image, D: merged image) and TiO2 coating (F: DAPI image, G: Cy3 image, H: FITC image, I: merged image) in osteogenic medium exhibited a more intense fluorescence of Cy3 and FITC compared with cells cultured in standard medium (E, J: merged images).

Article Snippet: After co-incubation with primary rabbit polyclonal anti-human osteopontin antibodies (1 : 100; Proteintech Group, Chicago, USA) and primary mouse monoclonal anti-human Runx2 antibodies (1 : 100; ProSci, Poway, USA) overnight at 48C, all the samples were then washed with PBS and co-incubated with Cy3-labeled goat anti-rabbit IgG (1:1000; Beyotime, Beijing, China) and fluorescein (FITC)-labeled goat anti-mouse IgG (1:1000; Beyotime) for 1 h at RT.

Techniques: Immunofluorescence, In Vitro, Labeling, Cell Culture, Fluorescence

Journal: PLoS ONE

Article Title: Demineralized bone matrix used for direct pulp capping in rats

doi: 10.1371/journal.pone.0172693

Figure Lengend Snippet: (A): Representative immunohistochemical images of the blank control group, the Ca(OH) 2 group and the DBM group. a: The blank control group. Pulp morphology was normal. b- f: Ca(OH) 2 group (1, 3, 7, 14, 28 days). g- k: DBM group (1, 3, 7, 14, 28 days). (B): Mean IOD value of Runx2. *means significant difference.

Article Snippet: The sections were deparaffinized with xylene, hydrated in a series of descending grades of ethanol, and then rinsed briefly with PBS for the primary antibodies; the sections were incubated overnight at 4°C with polyclonal anti- Runx2, COL I, OCN and DSP (Wuhan Boster Biological Technology, Wuhan, China).

Techniques: Immunohistochemical staining, Control

Journal: The Journal of Experimental Medicine

Article Title: The infarcted myocardium solicits GM-CSF for the detrimental oversupply of inflammatory leukocytes

doi: 10.1084/jem.20170689

Figure Lengend Snippet: GM-CSF is produced by cardiac fibroblasts after MI in mice and humans. (A) Csf2 mRNA levels in the infarcted tissue, uninfarcted tissue, spleen, BM, and mediastinal lymph node 1, 3, and 7 d after MI. Control mice were analyzed before MI (day 0; n = 3–5 per group from at least two independent experiments). *, P < 0.05, **, P < 0.01 vs. control. (B) Il-3 and Il-5 mRNA expression levels in infarcted tissue 1, 3, and 7 d after MI. Control mice were analyzed before MI (day 0; n = 5–6 per group from two independent experiments). (C) GM-CSF protein concentration in the heart homogenate of WT, Csf2 −/− , and Csf2rb −/− mice 1, 3, and 7 d after MI. Control mice were analyzed before MI (day 0; n = 3–5 per group from two independent experiments) *, P < 0.05, **, P < 0.01 vs. control. (D) Flow cytometric gating strategy to determine total leukocytes (Leuk), endothelial cells (ECs), fibroblasts (FBs), and other stromal cells (Other) in the heart. (E) Expression levels of genes encoding periostin ( Postn ), platelet-derived growth factor receptor-α ( Pdgfra ), and collagen 1a1 ( Col1a1 ) in infarcted tissue and indicated cells sorted from infarcted myocardium 1 d after MI. Values are normalized by gene levels in tissue ( n = 5–6 per group from two independent experiments). (F) Gene levels of Myh6 , which is specific for cardiomyocytes, in the heart tissue and the sorted heart subpopulations defined in D before and 1 d after MI. Data were normalized to 0 d tissue ( n = 5–6 per group from two independent experiments). (G) Csf2 mRNA expression levels in the sorted heart subpopulations defined in D before and 1 d after MI ( n = 4–5 mice from at least two independent experiments). *, P < 0.05 vs. 0 d. (H) Csf2 mRNA expression levels in infarcted tissue of WT, Myd88 −/− , Tlr2 −/− , Tlr4 −/− , Tlr7 −/− , Tlr9 −/− , Il1r −/− , and Ticam1 −/− (the gene that encodes TRIF) mice 1 d after MI ( n = 4–8 mice from two to three independent experiments). *, P < 0.05 vs. WT heart. (I) GM-CSF protein concentration in supernatants of cardiac fibroblasts cultured in the presence or absence of TLR3 or TLR9 agonists ( n = 4–6 per group from at least two independent experiments). (J) Representative immunohistochemical images of C4d staining in heart sections of patients with or without MI. Acute and late infarcts indicate the heart sections from patients who died within 2 d (acute) or 7 d (late) after MI onset. Bars, 1 mm. (K) Identification of GM-CSF + vimentin + cardiac fibroblasts (white arrows) in C4d + area indicated by the square in J. Bars, 20 µm. (L) Quantification of GM-CSF + cells in heart sections of patients with or without MI. Cells were counted in 25 randomly selected fields of view per sample. **, P < 0.01, ***, P < 0.001 vs. no MI; ## , P < 0.01 vs. vimentin − . For statistical analysis, two-tailed unpaired t test was performed to compare two groups, and one-way ANOVA followed by Tukey’s test was performed for multiple comparisons. Results are shown as mean ± SEM.

Article Snippet: Quantitative real-time TaqMan PCR was performed using the following TaqMan primers (Applied Biosystems): Il-1β (Mm01336189_m1), Il-3 (Mm00439631_m1), Il-5 (Mm00439646_m1), Il-6 (Mm00446190_m1), Mmp9 (Mm00442991_m1), Col1a1 (Mm00801666_g1), Pdgfra (Mm00440701_m1), Postn (Mm01284919_m1), Csf2 (Mm01290062_m1), Cxcl1 (Mm04207460_m1), Cxcl2 (Mm00436450_m1), Ccl2 (Mm00441242_m1), Ccl3 (Mm00441259_g1), Ccl5 (Mm01302427_m1), Ccl7 (Mm00443113_m1), Cx3cl1 (Mm00436454_m1), Ccl2 (Mm00441242_m1), Myh6 (Mm00440359_m1), Il-23a (Mm00518984_m1), and housekeeping gene Gapdh (Mm99999915_g1).

Techniques: Produced, Control, Expressing, Protein Concentration, Derivative Assay, Cell Culture, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN in M1 macrophage‐derived SFs mediates the formation of traumatic HO. A) High‐throughput sequencing was performed between SFs derived from macrophages and M1‐macrophages in vitro and between the sham group and the tendon lesions at 7 days in vivo. An intersection Venn diagram was drawn. B) WB analysis was used to detect the expression of POSTN proteins in Mφ‐SFs, M1‐SFs, and NFκB knock out groups, N = 3, ** p < 0.01, *** p < 0.001, **** p < 0.0001. C) IHC staining was used to detect the expression of POSTN in the sham, positive, and NFκB knock‐out groups, N = 6. D) Immunofluorescence staining for the Runx2, OCN, OPN for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 3. E) WB analysis was used to detect the expression of osteogenic‐related protein levels (Runx2, OCN, OPN) for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 3, **** p < 0.0001. F) ALP and ARS staining for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 6, **** p < 0.0001. G) Immunofluorescence staining for the Runx2 of tendons and Micro‐CT analysis of HO formation in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 6, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: Derivative Assay, Next-Generation Sequencing, In Vitro, In Vivo, Expressing, Knock-Out, Immunohistochemistry, Immunofluorescence, Staining, Micro-CT

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN promotes the formation of traumatic HO by enhancing β‐oxidation of fatty acids. A) High‐throughput whole‐transcriptome sequencing was performed and showed by Reactome pathways enrichment analysis of RNA‐seq data between the sham group and tendon lesions at 7 days, N = 3 B) Immunofluorescence staining for LCAD and MCAD of TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups. C) WB analysis was used to detect the expression of LCAD and MCAD proteins for TDSCs in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 3, **** p < 0.0001. D) Seahorse test was used to detect the oxidative phosphorylation level in the osteogenic induced TDSCs in addition of M1‐SFs or M1‐SFs with POSTN knockout groups, N = 3, ● represented M1‐SFs with POSTN knockout groups and ▲ represented M1‐SFs groups. E) Fluorescence and light microscope and WB analysis were used to confirm the success of downregulation of LCAD transfection for TDSCs, N = 3. F) Immunofluorescence staining for the LCAD and MCAD (red), co‐localized with PDGFRα(green) of tendons in addition of M1‐SFs with POSTN knockout groups, with or without sh‐LCAD, N = 3. G) WB analysis was used to detect the expression of LCAD and MCAD of TDSCs in addition of M1‐SFs with POSTN knockout groups, with or without sh‐LCAD, N = 3, **** p < 0.0001. H) ALP and ARS staining were used to detect the osteogenesis of TDSCs in addition of M1‐SFs with POSTN knockout groups, with or without sh‐LCAD, N = 6, **** p < 0.0001. I) Immunofluorescence staining for the Runx2 of tendons and micro‐CT analysis of HO formation in the M1‐SFs and M1‐SFs with POSTN knockout groups, N = 6, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: High Throughput Screening Assay, Sequencing, RNA Sequencing, Immunofluorescence, Staining, Knock-Out, Expressing, Phospho-proteomics, Fluorescence, Light Microscopy, Transfection, Micro-CT

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN enhances osteogenic propensity by binding to PTK7. A) Mass spectrometry analysis was used and presented through a Venn diagram to detect the potential molecules that showed increased binding to POSTN in the disease model between the sham group and the tendon injury group at 7 days, N = 3. B) Mass spectrum of the binding between PTK7 and POSTN. C) According to the Score Sequest HT, PTK7 ranks first among the molecules that bind to POSTN in the heterotopic ossification model. D) Docking images showing the predicted binding position of POSTN and PTK7 protein. E) IF staining showed co‐localization of POSTN and PTK7 in the cytoplasm of TDSCs, N = 3. F) IF staining showed co‐localization of POSTN and PTK7 in the heterotopic ossified tissue, N = 6. G) Co‐IP analysis was used to verified the bind relationship between POSTN and PTK7 in the osteogenic induced TDSCs, N = 3. H) Co‐IP analysis was used to verified the bind relationship between POSTN and PTK7 in the heterotopic ossified tissue, N = 3. I) Immunofluorescence staining indicated that overexpression of PTK7 could upregulate the protein expression levels of LCAD and MCAD in the absence of POSTN in vitro, N = 3. J) ALP and ARS staining was used to detect the osteogenesis of TDSCs in the POSTN‐/‐ and POSTN‐/‐&OV‐PTK7 groups, N = 6, **** p < 0.0001. K) Immunofluorescence staining for the Runx2 of tendons and micro‐CT analysis of HO formation in the POSTN‐/‐ and POSTN‐/‐&OV‐PTK7 groups, N = 6, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: Binding Assay, Mass Spectrometry, Staining, Co-Immunoprecipitation Assay, Immunofluorescence, Over Expression, Expressing, In Vitro, Micro-CT

Journal: Advanced Science

Article Title: POSTN‐Mediated Interplay of M1 Polarized Macrophage with Tendon‐Derived Stem Cells to Drive Traumatic Heterotopic Ossification Formation through PTK7/ATK Signaling?

doi: 10.1002/advs.202507951

Figure Lengend Snippet: POSTN promotes the osteogenic transition of TDSCs by mediating the phosphorylation of AKT at the S124 site. A) High‐throughput sequencing was performed between TDSCs treated with SFs derived from M1 macrophages and SFs derived from M1 macrophages with the POSTN protein knocked out respectively and between overexpressed phosphorylation at the AKT S124 site and the mutated S124 site respectively. An intersection Venn diagram was drawn. B) WB analysis was used to detect the expression of CPT1 proteins of TDSCs in the M1‐SFs, POSTN‐/‐, sh‐PTK7, and mut‐S124 groups, N = 3, **** p < 0.0001. C) Immunofluorescence staining for LCAD and MCAD of TDSCs in the POSTN‐/‐, POSTN‐/‐&sh‐CPT1 and mut‐S124&sh‐CPT1 groups, N = 3. D) ALP and ARS staining were used to detect the osteogenesis of TDSCs in the POSTN‐/‐, POSTN‐/‐&sh‐CPT1 and mut‐S124&sh‐CPT1 groups, N = 6, **** p < 0.0001. E) WB analysis was used to detect the expression of LCAD and MCAD proteins of TDSCs in the POSTN‐/‐, POSTN‐/‐&sh‐CPT1 and mut‐S124&sh‐CPT1 groups, N = 3, **** p < 0.0001.

Article Snippet: The recombinant POSTN protein and its inhibitor HY‐RS16974 were purchased from MedChemExpress (Shanghai, China).

Techniques: Phospho-proteomics, Next-Generation Sequencing, Derivative Assay, Expressing, Immunofluorescence, Staining

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