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Contributor Information

  • Name Sharon Tooze
  • Institute Cancer Research UK, London Research Institute: Lincoln's Inn Fields

Tool Details

  • Tool name: HEK293A EGFP-LC3 Cell Line
  • Tool type: Cell Lines
  • Tool sub-type: Continuous
  • Parental cell line: HEK293
  • Organism: Human
  • Tissue: Embryo
  • Disease: Cancer; Neurodegeneration; Infectious disease
  • Growth properties: Autophagy
  • Model: Transgenic
  • Conditional: Yes
  • 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.
  • Research area: Cancer; Apoptosis and autophagy; Cell signaling and signal transduction; Cell biology; Drug development; Metabolism
  • 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).
  • Cellosaurus ID: CVCL_W347
  • Additional notes: HEK293 cells were obtained under licence from AdVec

  • For Research Use Only

Target Details

  • Target: GFP-LC3B (Rat)

Application Details

  • Application notes: Stable cell line expressing EGFP-tagged LC3 (rat) in background of HEK293A cell line.

Handling

  • Format: Frozen
  • 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).
  • Shipping conditions: Dry ice

Documentation

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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.