Journal: Endocrinology

Article Title: Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.

doi: 10.1210/endo.140.10.7038

Figure Lengend Snippet: FIG. 8. Stat6 activation by IL-4. Normal human prostate epithelial cells (PrEC) in primary culture, LnCAP and PC-3 prostate cancer cells, ZR-75–1 and BT-20 breast cancer cells, HaCaT human immor- talized keratinocytes, HT-29 and Caco-2 human colon cancer cells, ME-180 human cervix cancer cells or JAR and JEG-3 human cho- riocarcinoma cells were incubated in the presence of absence of IL-4 (10 ng/ml for 30 min). Analysis of Stat6 activation using EMSA was performed as described in Materials and Methods using a well es- tablished Stat6 responsive element derived from the IgE-promoter. A Stat6 antibody was included in the binding reaction where indicated.

Article Snippet: The membrane was probed with a Stat6 antibody (Santa Cruz Biotechnology, Inc., Santa Clara, CA, catalog no. SC-621X).

Techniques: Activation Assay, Incubation, Derivative Assay, Binding Assay

Journal: Endocrinology

Article Title: Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.

doi: 10.1210/endo.140.10.7038

Figure Lengend Snippet: FIG. 7. Stat6 expression in cell types derived from peripheral tissues. Ten micrograms of total cell extract from ZR-75–1 and BT-20 breast cancer cells, normal human prostate epithelial cells (PrEC) in pri- mary culture, LnCAP and PC-3 prostate cancer cells, HaCaT human immortalized keratinocytes; HT-29and Caco-2 human colon cancer cells, ME-180 human cervix cancer cells, JAR and JEG-3 human choriocarcinoma and 293 human embryonic kidney cells were sepa- rated on SDS-PAGE. Western blot analysis was performed as de- scribed in Materials and Methods. Equal sample loading and transfer efficiency was confirmed by staining of the membrane with Ponceau Red.

Article Snippet: The membrane was probed with a Stat6 antibody (Santa Cruz Biotechnology, Inc., Santa Clara, CA, catalog no. SC-621X).

Techniques: Expressing, Derivative Assay, SDS Page, Western Blot, Staining, Membrane

Journal: Endocrinology

Article Title: Induction of 3beta-hydroxysteroid dehydrogenase/isomerase type 1 expression by interleukin-4 in human normal prostate epithelial cells, immortalized keratinocytes, colon, and cervix cancer cell lines.

doi: 10.1210/endo.140.10.7038

Figure Lengend Snippet: FIG. 9. IL-4-activated Stat6 binds to consensus Stat6 sequence sites in the 3b-HSD type 1 gene promoter. Normal human prostate epi- thelial cells (PrEC) in primary culture were incubated in the presence of absence of IL-4 (10 ng/ml for 30 min). Analysis of Stat6 activation using EMSA was performed as described in Materials and Methods using the 3b-HSD type 1 Stat6#1 and Stat6#2 probes from the 3b- HSD type gene promoter. A Stat6 antibody was included in the bind- ing reaction where indicated.

Article Snippet: The membrane was probed with a Stat6 antibody (Santa Cruz Biotechnology, Inc., Santa Clara, CA, catalog no. SC-621X).

Techniques: Sequencing, Incubation, Activation Assay

Journal: AIDS Research and Therapy

Article Title: The dynamic changes of interferon lambdas related genes and proteins in JAK/STAT pathway in both acute and chronic HIV-1 infected patients

doi: 10.1186/s12981-017-0158-7

Figure Lengend Snippet: Nucleotide sequences of the primers used for real-time PCR

Article Snippet: Antibodies (Abs) used in this study were as follows: mouse phycoerythin (PE)-conjugated anti-human IFN-alpha/beta R2 Ab (clone MMHAR-2, R&D Systems), mouse PE-conjugated anti-human IFN-lambdas R1 Ab (clone 601106, R&D Systems), mouse PE-conjugated anti-human IFN-gamma Receptor 1 (clone GIR-208, Thermo Fisher Scientific), Mouse eFluor ® 450-conjugated anti-human STAT1 (KIKSI0803, Thermo Fisher Scientific), Rabbit FITC anti-Human STAT2 (Thermo Fisher Scientific), Mouse PE-Cyanine7-conjugated anti-human STAT3 (clone LUVNKLA, Thermo Fisher Scientific), mouse APC-conjugated anti-human STAT4 (clone 4LURPLE, Thermo Fisher Scientific), mouse FITC-conjugated anti-human STAT5 (clone SRBCZX, Thermo Fisher Scientific), mouse PerCP-eFluor ® 710-conjugated anti-human STAT6 (clone CHI2S4N, Thermo Fisher Scientific).

Techniques: Sequencing

Journal: Cellular and Molecular Immunology

Article Title: IL-33 induces immunosuppressive neutrophils via a type 2 innate lymphoid cell/IL-13/STAT6 axis and protects the liver against injury in LCMV infection-induced viral hepatitis

doi: 10.1038/cmi.2017.147

Figure Lengend Snippet: IL-13/ILC2 induces Arg-1 expression of neutrophils through STAT6. (a) Neutrophils were stimulated with rIL-13 (40 ng/ml) or ILC2 supernatant for the indicated times. Anti-IL-13 neutralization Ab (2 μg/ml) was also used. The levels of p-STAT6 in neutrophils were analyzed by phosflow cytometry. (b) The mean fluorescence intensities (MFIs) are shown. (c) STAT6 inhibitor (100 nM) was used in the cell culture and the p-STAT6 was analyzed. (d) Neutrophils were cultured for 8 h with or without stimulation. The transcript levels of Arg-1 in neutrophils were measured by qPCR. The data are shown as the means±s.e.m., and the experiment was repeated independently three times. A two-tailed t-test was used to compare the two groups. One-way ANOVA was used to compare more than two groups. ***P<0.001. Ab, antibody; ANOVA, analysis of variance; IL, interleukin; ILC2, type 2 innate lymphoid cell; NS, no significance; qPCR, quantitative PCR; rIL-13, recombinant IL-13; STAT6, signal transducer and activator of transcription factor 6.

Article Snippet: The following phosflow Abs were purchased from BD Biosciences: Alexa Fluor 647 mouse anti-STAT3 (Py705), anti-STAT5 (pY694), and anti-STAT6 (pY641).

Techniques: Expressing, Neutralization, Cytometry, Fluorescence, Cell Culture, Two Tailed Test, Real-time Polymerase Chain Reaction, Recombinant

Journal: International Journal of Biological Sciences

Article Title: USP25 stabilizes STAT6 to promote IL-4-induced macrophage M2 polarization and fibrosis

doi: 10.7150/ijbs.99345

Figure Lengend Snippet: USP25 inhibit K48 ubiquitination of STAT6 to enhance STAT6/PPAR-γ signaling in macrophages. (A) Upper panel: representative Western blot results for STAT6, PPAR-γ and SOCS3 at different time points stimulated with IL-4. Lower panel: figures showing the data with three mice analyzed. (B) Western blot analysis of SOCS3 in the lung homogenates. (C) USP25 did not affect MAPK (p38 and ERK), Akt and PI3K signaling. (D) Upper panel: immunoprecipitation of proteins using USP25 antibody and immunoblotting analysis using STAT6 antibody. Lower panel: immunoprecipitation of proteins using STAT6 antibody and immunoblotting analysis using USP25 antibody. (E) BMDMs of WT and USP25 -/- were treated with MG132 (10 μM) for 6 h before lysis, followed by stimulation of IL-4 (10 ng/mL) for 30 min. Proteins were immunoprecipitated with STAT6 antibody and analyzed by immunoblotting with the indicated antibodies. (F) NIH/3T3 cells were transfected with plasmids encoding the indicated constructs. Proteins were immunoprecipitated with anti-STAT6 antibody and analyzed by immunoblotting with the indicated antibodies. IP, immunoprecipitation; STAT6, signal transducer and activator of transcription 6; PPAR-γ, peroxisome proliferator-activated receptor gamma; SOCS3, suppressor of cytokine signaling 3. *p < 0.05 and **p < 0.01.

Article Snippet: Primary antibodies include anti-STAT6 antibody (Cat:380957, ZENBIO, China), anti PPAR-γ antibody (Cat: A11183, ABCLONAL, China), anti-USP25 antibody (Cat: ab187156, Abcam, USA), anti-Arg1 antibody (Cat: ab91279, Abcam, USA), anti-Ubi antibody (Cat: 134953, Abcam, USA), anti-Ub-k48 antibody (Cat: 140601, Abcam, USA).

Techniques: Ubiquitin Proteomics, Western Blot, Immunoprecipitation, Lysis, Transfection, Construct