HEK293A EGFP-LC3 Cell Line

HEK 293A EGFP-LC3 is a stable epithelial cell line expressing EGFP-tagged LC3 (rat) in a HEK293A background.

Contributors

Inventor Institute
Sharon Tooze Cancer Research UK, London Research Institute: Lincoln's Inn Fields
Cat. #: 151480
Tool sub type: Continuous
Unit size: 1×10^6 cells / vial
Organism: Human
Tissue: Embryo
Gender: Male
Model: Transgenic
Growth properties: Autophagy
Product description: HEK 293A EGFP-LC3 is a stable epithelial cell line expressing EGFP-tagged LC3 (rat) in a HEK293A background. This line functions as a reporter cell line to monitor the induction of autophagy after aminoacid starvation or rapamycin treatment as well as autophagosome formation. This cell line uses GFP-LC3 as a standard assay read out and can be evaluated by both immunofluorescence and biochemical methods.
Gender: Male
CRISPR: Yes
Conditional: Yes
Production details: EGFP-LC3 (rat) plasmid was transfected into HEK293A cells. The cells were selected with 800ug/ml geneticin. Single cell clones were selected and grown up. Several clones were tested and clone 2GL9 was taken as the best based on a number of parameters (inc. GFP-LC3 expression levels and responsiveness to amino acid deprivation).
Additional notes: HEK293 cells were obtained under licence from AdVec
Cellosaurus ID: CVCL_W347
Parental cell line: HEK293
Disease: Cancer;Cancer
Format: Frozen
Shipping conditions: Dry ice
Growth medium: DMEM + 10% FCS + 2mM Glutamine. (400ug/ml geneticin can be added for maintainance but this is not normally necessary as the GFP-LC3 expression level is maintained over at least 20 passages).
Recommended controls: HEK293-A parental line
Mycoplasma free: Yes
References:

Chan et al. 2007. J Biol Chem. 282(35):25464-74. PMID: 17595159.

siRNA screening of the kinome identifies ULK1 as a multidomain modulator of autophagy.

Kchl et al. 2006. Traffic. 7(2):129-45. PMID: 16420522.

Microtubules facilitate autophagosome formation and fusion of autophagosomes with endosomes.

Images

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HEK293A EGFP-LC3 Cell Line. Image courtesy of the European Collection of Authenticated Cell Cultures (ECACC)

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