Journal: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

Article Title: Modulation of mucus production by interleukin-13 receptor alpha2 in the human airway epithelium.

doi: 10.1111/j.1365-2222.2007.02871.x

Figure Lengend Snippet: Fig. 10. Western blot analysis of signal transducer and activator of transcription 6 (STAT6) (a) and the phosphorylated, active form of STAT6 (p-STAT6) (b) in the cell lysates of normal human bronchial epithelial cells incubated with IL-13 (10 ng/mL) or IL-4 (1 ng/mL) for 14 days. Lanes show the data from samples treated with either the vehicle of IL-13 or IL-4 (PBS) (lane 1), IL-13 (10 ng/mL) plus the vehicle of antibody (PBS) (lane 2), IL-13 plus anti-IL-13Ra1 antibody (10 mg/mL) (lane 3), IL-13 plus recombinant human soluble IL-13Ra2/Fc (rsIL-13Ra2, 4 ng/mL) (lane 4), IL-4 (1 ng/mL) plus the vehicle of antibody (PBS) (lane 5) or with IL-4 (1 ng/mL) plus rsIL-13Ra2 (4 ng/mL) (lane 6). The arrows show STAT6 (a) and p-STAT6 (b) (100120 kDa). Data are representative of three differ- ent experiments.

Article Snippet: For STAT6 and the p-STAT6, the membranes were then blocked with 5% skimmed milk in Tris-buffered saline containing 0.1% Tween 20 and incubated with either a monoclonal anti-human/ mouse STAT6 antibody (R&D Systems Inc.) or an affinitypurified rabbit anti-phospho-STAT6 (Y641) antibody (R&D Systems Inc.).

Techniques: Western Blot, Incubation, Recombinant

Journal: Molecular Cancer Therapeutics

Article Title: Opposing control of rhabdomyosarcoma growth and differentiation by myogenin and interleukin 4

doi: 10.1158/1535-7163.mct-08-0678

Figure Lengend Snippet: Figure 4. A, forced expression of myogenin inhibited RD/12 cell migra- tion and blocked directed migration toward IL-4. Columns, mean number of cells migrated in the lower compartment containing medium with or without IL-4, four independent replicates; bars, SE. Migration of RD/ 12-Myog cells was significantly lower than parental RD/12 cells (P < 0.001, Student's t test); IL-4 significantly (stars, P < 0.05, Student's t test) increased migration only of RD/12 and RD/12-Neo cells, but not of RD/12-Myog and RD/18 cells expressing high myogenin levels. IL-4 re- ceptor level (B) and signaling (C) in rhabdomyosarcoma cells with forced myogenin expression. Total proteins (100 μg) were loaded for the evalua- tion of total-STAT6, phospho-STAT6 (STAT6 P), IL-4 Rα, and IL-13 Rα ex- pression. Actin (15 μg of total proteins) was used as a housekeeping protein for sample normalization. M, marker of molecular weight.

Article Snippet: Mouse antihuman IL-13 Rα1 monoclonal antibody (clone 419718, 2 μg/mL), goat anti-human IL-4 Rα antibody (0.2 μg/mL), rabbit anti–phospho-STAT6 (Y641) antibody (0.5 μg/mL; R&D Systems), rabbit anti-total STAT6 (1.5 μg/mL, M-20; Santa Cruz Biotechnology), and rabbit anti-actin (Sigma) were used as primary antibodies.

Techniques: Expressing, Migration, Marker, Molecular Weight

Journal: Nutrients

Article Title: Asaronic Acid Inhibited Glucose-Triggered M2-Phenotype Shift Through Disrupting the Formation of Coordinated Signaling of IL-4Rα-Tyk2-STAT6 and GLUT1-Akt-mTOR-AMPK

doi: 10.3390/nu12072006

Figure Lengend Snippet: ELISA data showing temporal response and upregulation of IL-10 secretion ( A , B ) and Western blot data showing upregulation of expression of arginase-1, CD163, PPARγ, phospho-Tyk2, and phospho-STAT6 by asaronic acid ( C – E ). J774A.1 macrophages were exposed to 40 ng/mL IL-4 in the absence and presence of 1–20 μM asaronic acid for up to 48 h. For the measurements of IL-10 secretion, IL-10 in cell culture media was detected by using an ELISA kit ( A , B ). Cell lysates were subject to 8–12% SDS-PAGE and Western blot analysis with a primary antibody against arginase-1, CD163, PPARγ, phospho-Tyk2, and phospho-STAT6. β-Actin antibody was used as an internal control. The bar graphs (mean ± SEM, n = 3) in the bottom panels represent quantitative results of the upper blot bands obtained from a densitometer. Mean values in respective bar graphs not sharing a same lower-case alphabet letter are significantly different at p < 0.05.

Article Snippet: Cells were treated with 5% BSA for 1 h. For the Immunofluorescent cytochemical staining, cells were incubated with a specific primary antibody against TGF-β or phospho-STAT6 overnight and further with Cy3-conjugated or FITC-conjugated IgG for 1 h (Rockland, Pottstown, PA, USA) and washed with phosphate-buffered saline-tween 20.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Cell Culture, SDS Page, Control

Journal: Nutrients

Article Title: Asaronic Acid Inhibited Glucose-Triggered M2-Phenotype Shift Through Disrupting the Formation of Coordinated Signaling of IL-4Rα-Tyk2-STAT6 and GLUT1-Akt-mTOR-AMPK

doi: 10.3390/nu12072006

Figure Lengend Snippet: Blockade of induction of phospho-Tyk2 and phospho-STAT6 by asaronic acid. J774A.1 macrophages were exposed to 33 mM glucose in the absence and presence of 1–20 μM asaronic acid for 48 h. Cell lysates were subject to 8–12% SDS-PAGE and Western blot analysis with a primary antibody against phospho-Tyk2 ( A ). β-Actin antibody was used as an internal control. The bar graphs (mean ± SEM, n = 3) in the panels represent quantitative results of the left blot bands obtained from a densitometer. Mean values in respective bar graphs not sharing a same lower-case alphabet letter are significantly different at p < 0.05. Immunocytochemical analysis showing phospho-STAT6 induction in glucose-loaded macrophages ( B ). The phospho-STAT6 localization was confirmed by FITC-green staining in macrophages exposed to 33 mM glucose ( n = 3). Nuclear staining was done with DAPI (blue). Magnification: 200-fold.

Article Snippet: Cells were treated with 5% BSA for 1 h. For the Immunofluorescent cytochemical staining, cells were incubated with a specific primary antibody against TGF-β or phospho-STAT6 overnight and further with Cy3-conjugated or FITC-conjugated IgG for 1 h (Rockland, Pottstown, PA, USA) and washed with phosphate-buffered saline-tween 20.

Techniques: SDS Page, Western Blot, Control, Staining