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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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PBL Assay blocking ab for the type i ifn receptor (clone mmhar-2) antibody
Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.
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Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Functional Effects of TGF-beta1 on Mesenchymal Stem Cell Mobilization in Cockroach Allergen Induced Asthma

doi: 10.4049/jimmunol.1303461

Figure Lengend Snippet: Epithelium-conditioned medium (ECM) induces MSC migration. (A) Air-liquid interface (ALI) culture system for airway epithelial cells. (B) Transwell assays for migration of MSCs. (C) Number of migrated cells in (B). (D) Levels of active TGF-β1 in ECM from un-challenged and challenged epithelial cells, n=3. (E) Number of migrated cells after ECM was treated with TGF-β1 neutralizing antibody (TGF-β1 Ab) or control antibody (IgG). (F) Number of migrated cells after ECM was treated with different doses of TβR1 inhibitor (SB505124). (G-H) Number of migrated cells after MSCs was directly treated with the addition of different doses of recombinant TGF-β1 into DEME in the lower chamber of Transwell (See B). Numbers of migrated cells were counted in five views (×20). Bars (C, E, and F) represent mean ± SEM of 3 independent experiments. *P<0.05 **P<0.01. BEC: bronchial epithelial cell.

Article Snippet: Briefly, BALF cells were first blocked with 10μg/ml IgG Fc receptor blocking reagent(2.4G2, BD Biosystems) for 10min, then stained with the mixture of the following antibodies, Anti-CD107b (Mac-3) FITC(M3/84,BD Biosystems); anti-Siglec-F-PE (E-50-2440, BD Biosystems); Anti-Ly-6G (Gr-1) PerCP-Cyanine5.5 (RB6-8C5, eBioscience); anti-CD3ε APC(145-2C11, eBioscience) and anti-CD19 APC(1D3, eBioscience) for 30 min.

Techniques: Migration, Recombinant