KC-5526

293T-ELK1-KLB-Cell-Line

58458

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Background of 293T-ELK1-KLB-Cell-Line

ELK1 (ETS Like-1 protein) is an important transcription factor gene and a member of the ETS family. Transcription factors are proteins that can bind to specific sequences of DNA, thereby regulating the 'on' or 'off' states of other genes. ELK1 mainly participates in regulating key life processes such as cell proliferation, differentiation, survival, and apoptosis, particularly playing a core role in responding to extracellular signals (such as growth factors and stress signals).
Beta klotho (KLB) is a fundamental component in fibroblast growth factor receptor (FGFR) signaling as it serves as an obligatory coreceptor for the endocrine hormones fibroblast growth factor 19 (FGF19) and fibroblast growth factor 21 (FGF21). Through the development of FGF19 and FGF21 mimetics, KLB has emerged as a promising drug target for treating various metabolic diseases, such as type 2 diabetes (T2D), non-alcoholic fatty liver disease (NAFLD), and cardiovascular disease. As a non-canonical fibroblast growth factor, fibroblast growth factor 21 (FGF21) functions as an endocrine hormone that signals to distinct targets throughout the body. Interest in therapeutic applications for FGF21 was initially sparked by its ability to correct metabolic dysfunction and decrease body weight associated with diabetes and obesity.

Specifications

Catalog Number	KC-5526
Cell Line Name	293T-ELK1-KLB-Cell-Line
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous ELK1-KLB gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-ELK1-KLB-cell-line was generated using a lentiviral vector expressing the ELK1-KLB sequence.

Characterization

Figure 1: Characterization of ELK1-KLB overexpression in the 293T-ELK1-KLB stable clone using qPCR.

Figure 2: Characterization of ELK1-KLB in the 293T-ELK1-KLB stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% 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. Aaldijk AS, Verzijl CRC, Jonker JW, Struik D. Biological and pharmacological functions of the FGF19- and FGF21-coreceptor beta klotho. Front Endocrinol (Lausanne). 2023 May 16; 14: 1150222. doi: 10.3389/fendo.2023.1150222. 2.Flippo KH, Potthoff MJ. Metabolic Messengers: FGF21. Nat Metab. 2021 Mar;3(3):309-317. doi: 10.1038/s42255-021-00354-2.
2.Zhang Y, Gao L, Liu L, Chen H. ELK1 modulates SF3B3 transcriptional activity to stimulate proliferation and inhibit apoptosis in gastric cancer through the activation of the MAPK pathway. Neoplasma. 2025 Aug;72(4):275-285. doi: 10.4149/neo_2025_250219N83. PMID: 40958521.
3. Kalampounias G, Androutsopoulou T, Katsoris P. Mechanistic Insights and Clinical Implications of ELK1 in Solid Tumors: A Narrative Review. Cells. 2025 Aug 14;14(16):1257. doi: 10.3390/cells14161257. PMID: 40862737; PMCID: PMC12384596.