transcription stat 6  (Santa Cruz Biotechnology)

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits M1 macrophage polarization and promotes M2 macrophage polarization in BMDM. (A) The cytotoxicity of Sauchinone on BMDM was detected by CCK-8 kit. (B-G) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Tnf-α , Il-6 , Il −12 , Inos , Il-1β , Cox-2 and Nlrp3 (B) were detected by RT-qPCR. TNF-α, IL-6, IL-12 and NO levels in culture supernatants (C) were measured by ELISA or Griess assay. The protein levels of p-STAT-1, STAT-1, and INOS in cell lysates (D) were detected by western blot. The percentages of INOS + , MHCⅡ + and CD86 + BMDM (E) were measured by FACS. (F-H) BMDM were stimulated with IL-4 plus IL-13 to induce M2 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Arg-1, Fizz1 , Ym-1 , Cd206 , Cd301 , and Dectin-1 (F) were detected by RT-qPCR. The protein levels of p-STAT-6 and STAT-6 in cell lysates (G) were detected by western blot. The percentages of CD301 + BMDM (H) were measured by FACS. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: CCK-8 Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Griess Assay, Western Blot

Journal: Clinical and molecular allergy : CMA

Article Title: Research Upregulation of CD23 (FcεRII) Expression in Human Airway Smooth Muscle Cells (huASMC) in Response to IL-4, GM-CSF, and IL-4/GM-CSF

doi: 10.1186/1476-7961-3-6

Figure Lengend Snippet: IL-4Rα and IL-2Rγc Expression in huASMC . A: IL-4Rα and IL-2Rγc expression in huASMC at baseline. Unstarved huASMC lysates were subjected to western blot analysis for IL-4Rα and IL-2Rγc using polyclonal anti-IL-4Rα (1:500 dilution, Santa Cruz), or monoclonal anti-IL-2Rγc (1:125 dilution, R&D Systems, Inc.). The nitrocellulose membranes were incubated with a 1:1,000 dilution of anti-rabbit or anti-mouse horseradish peroxidase linked whole antibody (Amersham). The immunoreactive protein bands were detected by ECL (Amersham). IL-2Rγc is minimally expressed in huASMC while IL-4Rα is expressed abundantly in huASMC.

Article Snippet: Standard Western blot analyses were performed to detect anti-STAT6 (1:500, Calbiochem, San Diego, CA) polyclonal rabbit, anti-p-STAT-6 (1:500, Calbiochem) polyclonal rabbit.

Techniques: Expressing, Western Blot, Incubation

Journal: Clinical and molecular allergy : CMA

Article Title: Research Upregulation of CD23 (FcεRII) Expression in Human Airway Smooth Muscle Cells (huASMC) in Response to IL-4, GM-CSF, and IL-4/GM-CSF

doi: 10.1186/1476-7961-3-6

Figure Lengend Snippet: Upregulation of IL-2Rγc Expression in huASMC by IL-4 and IL-4/GM-CSF . Alpha-smooth muscle isoactin positive Human ASMC (Clonetics) in T-75 flasks were starved for 24 h in 0.1% FBS containing medium M199. The cells were then either stimulated with BSA (vehicle) (1 μg/ml), IL-4 (0.5 nM), GM-CSF (0.4 nM), or IL-4/GM-CSF (0.5 nM/0.4 nM) for 24 hours. The IL-4 and IL-4/GM-CSF stimulated cells had increased IL-2Rγc expression compared to the BSA (vehicle) group. Fibronectin polyclonal rabbit antibody (Sigma) (1:250) was used as an irrelevant isotype control.

Article Snippet: Standard Western blot analyses were performed to detect anti-STAT6 (1:500, Calbiochem, San Diego, CA) polyclonal rabbit, anti-p-STAT-6 (1:500, Calbiochem) polyclonal rabbit.

Techniques: Expressing

Journal: Clinical and molecular allergy : CMA

Article Title: Research Upregulation of CD23 (FcεRII) Expression in Human Airway Smooth Muscle Cells (huASMC) in Response to IL-4, GM-CSF, and IL-4/GM-CSF

doi: 10.1186/1476-7961-3-6

Figure Lengend Snippet: Phosphorylation of STAT-6 by IL-4 and IL-4/GM-CSF in huASMC . Starved huASMC were incubated with either BSA vehicle control (1 μg/ml), IL-4 (0.5 nM), GM-CSF (0.4 nM), or IL-4/GM-CSF (0.5 nM/0.4 nM) for 15 minutes. Standard Western blot analyses were performed to detect STAT-6 and phosphorylated-STAT-6 (p-STAT-6) using a anti-STAT-6 polyclonal rabbit antibody (Calbiochem) and anti-p-STAT-6 polyclonal rabbit antibody (Calbiochem). Anti-rabbit horseradish peroxidase linked antibody was used as the secondary antibody. Protein bands were detected by ECL. STAT-6 was abundantly expressed by all four groups, while p-STAT-6 was only expressed in the IL-4 and IL-4/GM-CSF groups. Fibronectin polyclonal rabbit antibody (Sigma) was used as an irrelevant isotype control and was abundantly expressed in all four groups.

Article Snippet: Standard Western blot analyses were performed to detect anti-STAT6 (1:500, Calbiochem, San Diego, CA) polyclonal rabbit, anti-p-STAT-6 (1:500, Calbiochem) polyclonal rabbit.

Techniques: Incubation, Western Blot