The largest database of trusted experimental protocols
Sourced in United States, United Kingdom

STAT6 is a protein that plays a critical role in the regulation of gene expression. It is a member of the STAT (Signal Transducer and Activator of Transcription) family of transcription factors. STAT6 is essential for the differentiation and function of T helper 2 (Th2) cells, which are involved in the immune response to extracellular parasites and in the pathogenesis of allergic diseases.

Automatically generated - may contain errors

28 protocols using stat6

1

MUC5AC Expression Regulation in Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
A Lyn-specific siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). The antibodies for histology were purchased from Santa Cruz Biotechnology: MUC5AC (mucin 5AC, Clone # K-20; Cat # sc-16903), STAT6 (Clone # S-20, Cat # sc-621), β-actin (Clone # N-21, Cat # sc-130656), and phospho-STAT6 (Clone # Tyr641, Cat # sc-11762). The ELISA kits for IL-4, and IL-13 were purchased from Beijing Yonghui biotechnology Co. Ltd (Beijing, China). The MUC5AC-pGL3 luciferase vector was constructed by our laboratory based on the pGL3 luciferase reporter vector (Promega) following the manufacturer’s instructions.
+ Open protocol
+ Expand
2

Macrophage Polarization Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fluvastatin capsules were purchased from Novartis Pharmaceutical Co., Ltd. (Beijing, China). A Masson staining kit was purchased from Leagene Biotechnology Co., Ltd. (Beijing, China). A hematoxylin-eosin staining kit was purchased from Servicebio Technology Co., Ltd. (Wuhan, China). Cell Counting Kit-8 (CCK-8) was obtained from Fcmacs Biotech Co., Ltd. (Nanjing, China). A BCA protein assay kit was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Rosiglitazone (RSG, selective PPAR-γ agonist) and T0070907 (selective PPAR-γ antagonist) were obtained from Selleck Chemicals (Houston, USA). LPS was purchased from Sigma Chemical (St. Louis, USA). Recombinant rat IL-4 and INF-γ were purchased from PeproTech Inc. (New Jersey, USA). ELISA kits (IL-1, IL-6, TNF-α, IL4, IL-10, IL-13, and TGF-β1) were obtained from JinYiBai Biological Technology (Nanjing, China). The antibodies (JAK2, STAT6, and p-STAT1) were purchased from Santa Cruz Biotechnology (California, USA). The antibodies (PPAR-γ, STAT1, and p-PPAR-γ) were purchased from Bioss (Beijing, China). The antibodies (SOCS1, SOCS3, p-JAK2, and p-STAT6) were purchased from Affinity Biosciences. APC-anti-mouse CD86 was purchased from BioLegend (San Diego, USA). PE-Cy7-anti-mouse CD206 and Intracellular Fixation & Permeabilization set were purchased from Thermo Fisher Scientific (Massachusetts, USA).
+ Open protocol
+ Expand
3

Quantifying Nuclear Translocation of p65 and STAT6

Check if the same lab product or an alternative is used in the 5 most similar protocols
Nuclear extracts were prepared using a nuclear extract kit (Active Motif). Protein content was measured using Bradford reagent (Pierce, Rockford, IL). Immunoblotting was performed using antibodies to p65, STAT6, and PARP (Santa Cruz Biotechnology). Nuclear translocation of p65 and STAT6 was quantified using the ImageJ program from the NIH and normalized to the nucleus specific protein PARP.
+ Open protocol
+ Expand
4

Immunoblotting of EMT Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Polyclonal antibodies against p-Akt (Ser473), Akt, and p-GSK3β, and monoclonal antibodies to β-catenin and Vimentin, were purchased from Cell Signaling Technology (Danvers, MA, USA). Polyclonal antibodies to JNK, Slug, Sox2, p-Stat6, Stat6, Ang-2, and VEGF, and monoclonal antibodies to p-JNK, β-actin, and Oct4 were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Polyclonal antibody to Snail was purchased from Novus Biologicals (Littleton, CO, USA). The monoclonal antibody E-cadherin was obtained from BD Transduction Laboratories (San Jose, CA, USA). The polyclonal antibody Twist was purchased from Abcam Inc. (Cambridge, MA, USA). Monoclonal antibodies to IL-4 and IL-4Rα were obtained from R&D systems (Minneapolis, MN, USA). Anti-mouse and anti-rabbit Alexa Fluor 488 and 555-conjugated secondary antibodies were purchased from Thermo Fisher scientific (Waltham, MA, USA). Inhibitors of JAK, PI3K (LY294002), and JNK (SP600125) were purchased from Merck Millipore (Darmstadt, Germany). Recombinant IL-4 was obtained from R&D systems (Minneapolis, MN, USA).
+ Open protocol
+ Expand
5

Adipose Tissue Protein Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Hypothalami, epididymal fats, and liver tissues (n = 6/group) were isolated, and proteins extracted using the commercial Tissue Protein Extraction Reagents (T-PER, Pierce Biotechnology, Rockford, IL, USA). Equal amounts of proteins were resolved by SDS-PAGE, before being transferred to the polyvinylidine fluoride membrane. The membranes were blocked with 5% nonfat milk and incubated with primary antibodies, horseradish peroxidase-conjugated IgG, and enhanced chemiluminescence Western blotting reagents. The visualized signals were quantitated by a densitor meter. Primary antibodies used were: Akt, phospho-Akt, signal transducers and activators of transcription 3 (STAT3), phospho-STAT3, STAT6, phospho-STAT6, PRDM16, PGC-1α, UCP-1, CD68, CD206, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
+ Open protocol
+ Expand
6

Protein Expression Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues/cell lysate pellets were subjected to SDS-PAGE, transferred into 0.45 μm PVDF membranes and blocked 30 minutes at room temperature with 3% skim milk (Sigma-Aldrich), prior overnight incubation with the following primary antibodies: phospho-ATM S1981 and STAT6 (all from Santa Cruz Biotechnology, Dallas, TX); β-actin (Sigma-Aldrich); Vimentin, p-STAT6 Y641, Histone H3, HP1γ, and H3K27me3 (all from Abcam, Cambridge, UK); BAX, IRE1-α, phospho-EIF2A, phospho-CHK2 T387, CHOP, phospho-STAT3 Y705, and phospho-Histone H2AX S139 (all from Cell Signaling Technology); HP1β (Thermo Fischer Scientific, Waltham, MA); STAT3, BCL-2, BCL-XL, and E-cadherin (BD Biosciences, Franklin Lakes, NJ); phospho-PERK (US Biologicals, Salem, MA). After 30 min incubation with their respectively HRP-conjugated secondary antibodies (GE healthcare, Chicago, IL) membranes were developed using the SuperSignal West Pico Chemoluminescence Substrate (Thermo Fischer Scientific). Western blots were quantified using ImageJ 1.43u Software (National Institutes of Health, Bethesda, Maryland, USA. https://imagej.nih.gov/ij/, 1997–2016).
+ Open protocol
+ Expand
7

Signaling Pathways in B-cell Malignancies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ibrutinib was purchased from Selleckchem (Houston, TX, USA), cerdulatinib was provided by Portola Pharmaceuticals Inc. (South San Francisco, CA, USA), CpG (ODN2006, stimulatory CpG-ODN type B, human specific) was purchased from Invivogen (San Diego, CA, USA), IL-6 was from R&D Systems (Minneapolis, MN), and IL-4 and CD40L were from Enzo Life Sciences (Plymouth Meeting, PA, USA). Antibodies: anti-phosphorylated BTK (p-BTK) (Y223), p-IκBα (S32/36) p-STAT3 (Y705), STAT3, MCL-1, p-JAK1 (Y1022), p-JAK3 (Y980), p-STAT6 (Y641), p-STAT3 (S727), and p-JAK2 (T1007/1008) were purchased from Cell Signaling Technology (Danvers, MA, USA); anti-total BTK antibody, poly ADP-ribose polymerase (PARP), and the BrdU detection kit were from BD Biosciences (San Jose, CA, USA); anti-p65, STAT-6, STAT-3, JAK1, JAK2, JAK3 and GAPDH antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). For flow cytometry, FITC-anti-CD19 (clone HIB19) and PE-anti-CD5 (clone UCHT2) were purchased from eBioscience (San Diego, CA, USA). Alexa Fluor® 647-anti-p-AKT (Ser473) and Alexa Fluor® 488 anti-p-p44/42 MAPK (ERK1/2) (T202/Y204) were purchased from Cell Signaling Technology, and PE-anti-p-PLCγ2 (Y759) was purchased from BD Bioscience.
+ Open protocol
+ Expand
8

Multisite Retrospective Study of Rare Soft Tissue Sarcoma

Check if the same lab product or an alternative is used in the 5 most similar protocols
The study was approved by the Institutional Review Board (IRB) of each participating site. Candidate cases were retrieved from the pathology archives of Memorial Sloan Kettering Cancer Center (New York, NY, USA, n=8), Johns Hopkins University (Baltimore, MD, USA, n=3), University Medical Center Groningen (Groningen, Netherlands, n=1), and Mount Sinai Hospital (Toronto, ON, Canada, n=1). All cases were centrally reviewed by two pathologists (BX and CRA) to gather relevant pathologic parameters. A chart review was conducted at each individual site to collect demographic and outcome data. The 5-year and 10-year overall survival and disease-specific survival were calculated using SPSS software 24.0 (IBM Corporation, Armonk, NY, U.S.).
Immunohistochemistry studies were performed in a subset of cases using the following primary antibodies: CD99 (clone: O13, dilution 1:800, Labcorp, Princeton, NJ, USA), CD34 (clone: QBEnd-10, ready to use RTU, Ventana, Roche diagnostics, Oro Valley, AZ, UA), SATB2 (clone: EP281, dilution 1:00, Cell Marque corporation, Rocklin, CA, USA), and STAT6 (catalog number: SC-621, dilution 1:2500, Santa Cruz Biotechnology, Dallas, TX, USA).
+ Open protocol
+ Expand
9

Th2 Cell Differentiation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The splenocytes were isolated from sacrificed BALB/c mice. CD4+ T cells were isolated using anti-CD4 micro-beads according to the manufacturer’s instructions (Miltenyl Biotec., Auburn, CA). CD4+ T cells were stimulated with plate-bound anti-CD3 (0.5 ㎍/㎖) and anti-CD28 (1 ㎍/㎖). The cells were additionally treated with IL-2 (2 ng/ml) for the generation of Th0 or with IL-4 (10 ng/ml) and anti-IFNγ (5 ㎍/ml) for Th2. Three hours after initial culture, cells were treated with either vehicle or picroside II, and cultured for 3 days. Cell supernatants were used for ELISA, and cell pellets were harvested for real-time RT-PCR or western blot as described above.
SDS-PAGE was probed with antibodies against GATA3, phosphorylated STAT6 (Cell Signaling Technology, Danvers, MA) and STAT6 (Santa Cruz, Dallas, TX).
+ Open protocol
+ Expand
10

Comprehensive Immunohistochemistry Panel for Tumor Diagnosis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cases were identified in the consultation files of the authors. The tissue specimens were fixed in formalin and processed routinely for histopathology. Immunohistochemistry (IHC) was performed on 3-μm sections cut from paraffin blocks using a fully automated system (“Benchmark XT System”, Ventana Medical Systems Inc., 1910 Innovation Park Drive, Tucson, Arizona, USA) and the following antibodies: vimentin (V9, 1:100, Dako), keratin cocktail (clone AE1/AE3, 1:40, Zytomed, Berlin, Germany), p63 (SSI6, 1: 100, DCS), desmin (clone D33, 1:250, Dako), alpha smooth muscle actin (clone 1A4, 1:200, Dako), HMB45 (clone HMB45, 1:50, Enzo), Melan A (clone A103, 1:50, Dako), CD34 (clone BI-3C5, 1:200, Zytomed), ERG (EPR3864, prediluted, Ventana), CD31 (clone JC70A, 1:20, Dako), S100 protein (polyclonal, 1:2500, Dako), SOX10 (polyclonal, 1:25, DCS), PAX8 (polyclonal rabbit anti-PAX8, 1:50, Cell Marque), NSE (clone BBS/NC/VI-H1, 1:300, Dako), TTF1 (clone 8G7G3/1, 1:500, Zytomed Systems, Berlin, Germany), Napsin A (MRQ-60, ready-to-use, Medac), calretinin (polyclonal, 1:100, Zytomed), alpha-inhibin (clone R1, 1:50, Serotec), GFAP (Clone GFA, 1/1000, DakoPatts, Denmark), EMA (clone E29, 1:200, Dako), STAT6 (clone S-20, 1:1000, Santa Cruz Biotechnology), TFE3 (clone MRQ-37, 1:100, Cell Marque), Cathepsin-K (clone 3F9, 1:50, Abcam) and Ki67 (clone MiB1, 1:100, Dako).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!