Human STING (R232)-dependent STAT6 reporter cells
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Human STING (R232 variant)-dependent STAT6 HEK293 reporter cells
3-7 x 10e6 cells
STING-dependent STAT6 activation
in HEK-Blue™ STAT6-hSTING-R232 cells
Human STING-dependent STAT6 reporter HEK293 cells
HEK-Blue™ STAT6-hSTING-R232 cells are specifically designed to monitor the induction of STAT6-dependent signaling upon activation of STING. These cells were generated by stable overexpression of the most prevalent human (h)STING variant R232 , in a human embryonic kidney (HEK)293-derived cell line that expresses human STAT6 and a STAT6-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. STAT6-dependent SEAP activity is readily assessable in the supernatant using QUANTI-Blue™ Solution, a detection reagent.
STING (stimulator of interferon genes) is essential in the effective immune response against foreign or self-DNA through the sensing of cytoplasmic cyclic dinucleotides (CDNs) . The activation of STING causes a TANK-binding-kinase-I (TBK1)-dependent cascade, ultimately, leading to IFN regulatory factor (IRF3)-dependent type I IFN production and NF-κB-dependent inflammatory cytokine production . Additionally, signal transducer and activator of transcription 6 (STAT6) has been reported to be recruited to STING for TBK1-mediated phosphorylation during viral infection . Ultimately, the STING‑dependent activation of STAT6 induces a specific set of anti-viral chemokines responsible for immune cell homing, which leads to reduced viral replication . Notably, this specific 'viral' activation of STAT6 was found to be Janus kinase (JAK)-independent and is thus, distinct from the ‘canonical’ STAT6 pathway activated by IL-4 and IL-13, which is critical in adaptive immunity .
Features of HEK-Blue™ STAT6-hSTING-R232 cells:
- Verified overexpression of hSTING (PCR and functional assays)
- Functionally validated with a selection of STING ligands
- These cells do not respond to Type I IFNs
- Readily assessable STAT6-dependent SEAP reporter activity
- The stability for 20 passages has been verified
- Guaranteed mycoplasma-free
Applications for HEK-Blue™ STAT6-hSTING-R232 cells:
- Studying the role of STING-dependent STAT6 activation in response to viral infection
1. Yi, G. et al. 2013. Single nucleotide polymorphisms of human STING can affect innate immune response to cyclic dinucleotides. PLoS One 8, e77846.
2. Cheng, Z. et al. 2020. The interactions between cGAS-STING pathway and pathogens. Signal Transduct Target Ther 5, 91.
3. Chen, H. et al. 2011. Activation of STAT6 by STING is critical for antiviral innate immunity. Cell 147, 436-446.
STAT6 activation upon STING induction. HEK-Blue™ STAT6-hSTING-R232 and their parental cells (STAT6 reporter cells) were incubated with 2’3’-cGAMP (30 μg/ml), 2’3’-cGAM(PS)2 (Rp/SP) (ADU-S100; 30 μg/ml), cAIM(PS)2 Difluor (Rp/Sp) (30 μg/ml), 3’3’-cGAMP (30 μg/ml), or DMXAA (30 μg/ml) in cell culture medium. After overnight incubation, the STAT6 response was assessed by measuring the activity of SEAP in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are presented fold change over non-induced cells (mean ± SEM).
Antibiotic resistance: Blasticidin, Hygromycin B, and Zeocin®
Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine,10% (v/v) heat-inactivated fetal bovine serum (FBS), Pen‑Strep (100 U/ml-100 μg/ml), 100 μg/ml Normocin™
- STAT6-dependent SEAP reporter activity in response to various STING ligands and other cytokines has been validated.
- The stability for 20 passages, following thawing, has been verified.
- These cells are guaranteed mycoplasma-free.
This product is covered by a Limited Use License (See Terms and Conditions).Back to the top
- 3-7 x 106 HEK-Blue™ STAT6-hSTING-R232 cells in a cryovial or shipping flask
- 2 x 1 ml of HEK-Blue™ Selection (250x concentrate)
- 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.
- 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
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