Stat5 inhibitor

Manufactured by Merck Group

Sourced in United Kingdom, United States

STAT5 inhibitor is a laboratory research tool used to study the role of the STAT5 protein in cellular signaling pathways. It functions by selectively inhibiting the activity of the STAT5 transcription factor, which plays a key role in the regulation of gene expression. This product is intended for use in in vitro research applications to investigate the biological functions of STAT5 and its involvement in various cellular processes.

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Lab products found in correlation

Flagellin, by InvivoGen (1 mentions) Cl097, by InvivoGen (1 mentions) Il 12p40, by Thermo Fisher Scientific (1 mentions) Il 10, by BD (1 mentions) Cpg dna, by InvivoGen (1 mentions) Il 13, by BioLegend (1 mentions) Lipid a, by Peptide Institute (1 mentions) Il 22, by Thermo Fisher Scientific (1 mentions) Interleukin 4 (il 4), by BioLegend (1 mentions) Il 1β, by Thermo Fisher Scientific (1 mentions) Il 22, by R&D Systems (1 mentions) Il 10, by Thermo Fisher Scientific (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Tgf β, by BioLegend (1 mentions) Tgf β, by R&D Systems (1 mentions) Pam3cys, by Merck Group (1 mentions) Bay11 7082, by Merck Group (1 mentions) Bx795, by Merck Group (1 mentions) Sircol collagen assay kit, by Biocolor (1 mentions) Anti cd3, by Merck Group (1 mentions)

5 protocols using stat5 inhibitor

CD4+ T Cell Activation and STAT5 Inhibition

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Anti-CD3 (BD Biosciences; cat. no. 561798) and anti-CD28 (BD Biosciences; cat. no. 562764) antibodies were coated in each well on day 1, followed by addition of anti-IFN (BD Biosciences; cat. no. 551506) and anti-IL-4 (BD Biosciences; cat. no. 555090). Purified CD4⁺ T cells were seeded into a 24-well plate at a density of 1.5×10⁶ cell per well in a 1 ml volume. Subsequently, 10 ng/ml IL-7 (PeproTech, Inc.) and 100 µM STAT5 inhibitor (Merck KGaA) were added to the culture, and incubated for 72 h. The following culture groups were obtained: Negative control (NC) group, Anti-CD3 + anti-CD28 + DMSO; NC + IL-7 group, Anti-CD3 + anti-CD28 + IL-7 + DMSO; NA group, Anti-CD3 + anti-CD28 + DMSO; NA + IL-7 group, Anti-CD3 + anti-CD28 + IL-7 + DMSO; NA + IL-7 + STAT5 inhibitor group:, Anti-CD3 + anti-CD28 + IL-7 + STAT5 inhibitor. The cells in each group were detected by a flow cytometer (FACS Calibur; BD Biosciences) and analyzed using FlowJo 7.6.5 software (FlowJo LLC).

Zhang X., Zhang M., Jiang M, & Nong G. (2020). Effect of IL-7 on Th17 cell responses in a mouse model of neutrophilic asthma. Molecular Medicine Reports, 22(2), 1205-1212.

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Modulation of Human Macrophage Responses

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Human MDMs were treated with MDP (Bachem, Torrance, CA), lipid A (Peptides International, Louisville, KY), Pam3Cys (EMD Millipore, Billerica, MA), poly(I:C), flagellin, CL097, CpG DNA (Invivogen, San Diego, CA), 2ng/ml TGFβ, 10ng/ml IL4 (R&D Systems, Minneapolis, MN), 10ng/ml IL13 (Biolegend, San Diego, CA), 10ng/ml IL22, 10ng/ml IL10, or 10ng/ml TSLP (Peprotech, Rocky Hill, NJ) unless otherwise stated. In other cases cells were co-cultured with Salmonella enterica serovar Typhimurium at MOI 10:1. For inhibitor studies, cells were pre-incubated with 25μM STAT3 inhibitor peptide or 25μM STAT5 inhibitor (EMD Millipore) (unless otherwise indicated) for 1h before treatments. Supernatants were assayed for cytokine secretion per manufacturer instructions using antibodies to the following proteins: TNF, IL6, IL8, IL10 (BD Biosciences, San Jose, CA), IL12p40 or IL1β (eBioscience, San Diego, CA), TGFβ, IL4, IL13 (Biolegend), IL22 (R&D Systems), or TSLP (PeproTech, Rocky Hill, NJ).

Hedl M., Sun R., Huang C, & Abraham C. (2019). STAT3 and STAT5 signaling thresholds determine distinct regulation for innate-receptor-induced inflammatory cytokines and STAT3/STAT5 disease variants modulate these outcomes. Journal of immunology (Baltimore, Md. : 1950), 203(12), 3325-3338.

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Inhibitors Modulate IL-25 Signaling

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Human recombinant IL-25 was purchased from R&D System (Minneapolis, MN) and was dissolved in sterile 4 mM HCl containing at least 0.1% bovine serum albumin. U0126 (a specific inhibitor of ERK), SB203580 (a specific inhibitor of p38) and SP600125 (a specific inhibitor of JNK inhibitor for JNK1, JNK2 and JNK3) were obtained from Calibiochem (Billerica, MA). STAT5 inhibitor was purchased from Merck Millipore (Nottingham, United Kingdom). Bay 11–7082 (NF-κB inhibitor) and BX-795 (IRF3 inhibitor) were purchased from Sigma (St. Louis, MO). All inhibitors were dissolved in dimethyl sulfoxide. Sircol collagen assay kits were acquired from Biocolor Ltd (Belfast, N. Ireland, UK).

Park S.K., Jin Y.D., Park Y.K., Yeon S.H., Xu J., Han R.N., Rha K.S, & Kim Y.M. (2017). IL-25-induced activation of nasal fibroblast and its association with the remodeling of chronic rhinosinusitis with nasal polyposis. PLoS ONE, 12(8), e0181806.

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Modulating CD8+ T Cell Responses

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Purified CD8⁺ T cells were stimulated with recombinant human IL-7 (10 ng/mL; R&D Systems, Minneapolis, Minnesota, USA) [22 (link)] for 48 h in the presence of anti-CD3/CD28, along with either JAK inhibitor (10 µmol/L; Sigma-Aldrich, Temecula, California, USA), STAT5 inhibitor (250 µmol/L; Merck Millipore), or PI3K inhibitor (LY294002) (25 µmol/L; Sigma-Aldrich) as previously described [20 (link)]. Control cells were only stimulated with anti-CD3/CD28 for maintenance of CD8⁺ T cell survival. In certain experiments, stimulated CD8⁺ T cells were washed twice. 10⁴ of CD8⁺ T cells were co-cultured with 10⁵ of melanoma cell line SK-MEL-5 cells for 48 h.

He H., Qiao B., Guo S., Cui H., Zhang Z, & Qin J. (2022). Interleukin-7 regulates CD127 expression and promotes CD8+ T cell activity in patients with primary cutaneous melanoma. BMC Immunology, 23, 35.

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Shrimp Stat5 Inhibitor Viability Study

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Stat from shrimp is similar to human Stat5; therefore, Stat5 inhibitor from Merck was used to inhibit shrimp Stat. To ascertain the effect of Stat5 inhibitor on shrimp viability, the survival rate of shrimp after inhibitor injection was analyzed. Shrimp (8–10 g each individual, 20 shrimp per group) were divided into four groups, and injected with 1, 2 and 4 μg of Stat5 inhibitor, respectively. The dimethylsulfoxide (DMSO) injection group was used as the control. For the Co-IP assay, Stat5 inhibitor (2 μg) was injected into each shrimp, which were then challenged with V. anguillarum. DMSO injection was used as a control. The hemocyte proteins were collected for protein extraction 3 h after V. anguillarum challenge to further detect if Stat inactivation affected the interaction of βarr1 with Stat and TC45.

Sun J.J., Yang H.T., Niu G.J., Feng X.W., Lan J.F., Zhao X.F, & Wang J.X. (2016). β-Arrestin 1’s Interaction with TC45 Attenuates Stat signaling by dephosphorylating Stat to inhibit antimicrobial peptide expression. Scientific Reports, 6, 35808.

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