KC-6313

293T-STAT4-Luc2 Cell Line

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Home » 细胞系 » 293T-STAT4-Luc2 Cell Line

Background of 293T-STAT4-Luc2 Cell Line

The protein encoded by this gene is a member of the STAT family of transcription factors. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein is essential for mediating responses to IL12 in lymphocytes, and regulating the differentiation of T helper cells. Mutations in this gene may be associated with systemic lupus erythematosus and rheumatoid arthritis. Alternate splicing results in multiple transcript variants that encode the same protein. STAT4 (Signal Transducer And Activator Of Transcription 4) is a Protein Coding gene. Diseases associated with STAT4 include Disabling Pansclerotic Morphea Of Childhood and Systemic Lupus Erythematosus 11. Among its related pathways are Interleukin-12 family signaling and IL27-mediated signaling events. Gene Ontology (GO) annotations related to this gene include DNA-binding transcription factor activity and sequence-specific DNA binding.

Specifications

Catalog NumberKC-6313
Cell Line Name293T-STAT4-Luc2 Cell Line
Clone Number4B1
Host Cell Line293T
DescriptionStable 293T cell line expressing exogenous luciferase under the control of STAT4 responsive element.
QuantityTwo vials of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% DMEM + 20% FBS + 10% DMSO
Propagation MediumDMEM + 10% FBS + 75μg/mL Hygromycin B + 0.5μg/mL puromycin
Selection MarkerHygromycin B and Puromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:4-1:6 every 2-3 days; seed out at about 1 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative

Cell Line Generation

293T-STAT4-Luc2 Cell Line was generated using a lentiviral vector expressing the STAT4-Luc2 sequence.

Characterization

Figure 1: 293T-STAT4-Luc2 cell were seeded into 96-well plates, treated with IFN-γ in different concentrations for 16 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

  1. Prewarm culture medium (DMEM supplemented with 10% FBS, 75μg/mL Hygromycin B and 0.5μg/mL Puromycin) in a 37°C water bath.
  2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
  3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
  4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
  5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
  6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
  7. Incubate the flask at 37°C, 5% CO2 incubator.
  8. Split saturated culture 1:4-1:6 every 2-3 days; seed out at about 1 × 105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
  2. Keep the freezing medium on ice and label cryovials.
  3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
  4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
  5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
  6. Aliquot 1 mL of the cell suspension into each cryovial.
  7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
  8. Transfer vials to liquid nitrogen for long-term storage.

References

  1. Baghdassarian H, Blackstone SA, Clay OS, Philips R, Matthiasardottir B, Nehrebecky M, Hua VK, McVicar R, Liu Y, Tucker SM, Randazzo D, Deuitch N, Rosenzweig S, Mark A, Sasik R, Fisch KM, Pimpale Chavan P, Eren E, Watts NR, Ma CA, Gadina M, Schwartz DM, Sanyal A, Werner G, Murdock DR, Horita N, Chowdhury S, Dimmock D, Jepsen K, Remmers EF, Goldbach-Mansky R, Gahl WA, O'Shea JJ, Milner JD, Lewis NE, Chang J, Kastner DL, Torok K, Oda H, Putnam CD, Broderick L. Variant STAT4 and Response to Ruxolitinib in an Autoinflammatory Syndrome. N Engl J Med. 2023 Jun 15;388(24):2241-2252. doi: 10.1056/NEJMoa2202318. Epub 2023 May 31. PMID: 37256972; PMCID: PMC10392571.
  2. National Center for Biotechnology Information (2026). PubChem Pathway Summary for Pathway il12_stat4pathway, IL12 signaling mediated by STAT4, Source: Pathway Interaction Database. Retrieved February 4, 2026 from https://pubchem.ncbi.nlm.nih.gov/pathway/Pathway-Interaction-Database:il12_stat4pathway.
  3. Zinser E, Tan KL, Kim DS, O'Brien R, Winstanley A, Yong PFK. Differential Diagnosis: Hepatic Complications in Inborn Errors of Immunity. J Clin Med. 2023 Dec 3;12(23):7480. doi: 10.3390/jcm12237480. PMID: 38068532; PMCID: PMC10707526.
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