Cell line profile Ulk
Cell lines for autophagy research (see catalogue numbers below)
Cell line history
A group of mouse embryonic fibroblast cell lines have been deposited by Ximbio and are available from the Culture Collections Public Health England. These display different capabilities for uncoordinated (Unc)-51-like kinase 1 and 2 (UlkK1 and Ulk2) activity, which are known to play a critical role during the activation of autophagy.
ULK1/2 WT (SV40) - early log phase 24 hours ULK1/2 WT (SV40) – late log phase 72 hours
Key characteristics
Mouse embryonic fibroblast were derived from Ulk1 +/- and Ulk2 +/- knockout mice. These include single knockout cell lines for Ulk1 or Ulk2 or double knockout cell lines for both Ulk1 and Ulk2. The cell lines were immortalised either using a standard serial passaging protocol or the SV40 large T-antigen. There are also a pair of cell lines representing wild type phenotype for Ulk 1 and 2 which can be used as controls with the knockout Ulk cell lines.
Applications
Unc)-51-like kinase 1 and 2 (ULK1 and ULK2) activity are known to play a critical role during the activation of autophagy. Autophagy is the mechanism by which a cell degrades unnecessary or dysfunctional cellular components and has been implicated in several medical scenarios such as cancer, neurodegeneration and immunity related disorders.
Culture tips
The Ulk cell lines should be cultured in DMEM + 10% Foetal Calf Serum +2mM Glutamine. Trypsinize the cells with 0.05% Trypsin/EDTA and inoculate new cultures at 2 x 104 per cm2. Cultures must be incubated in a humidified 5% CO2/95% air incubator at 37◦C.
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Cell line name |
ECACC catalogue no. |
Description |
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MEF Ulk1 -/- Ulk2 -/- (DKO) (SV40) |
Mouse embryonic fibroblast, autophagy, Ulk1 -/- Ulk2 -/- double knockout SV40 immortalised |
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MEF Ulk1 -/- Ulk2 -/- (DKO) (SIM) |
Mouse embryonic fibroblast, autophagy, Ulk1 -/- Ulk2 -/- double knockout spontaneously immortalised |
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Ulk/1/2 WT MEF (SV40) |
Mouse embryonic fibroblast, autophagy, Unc51-like kinase SV40 immortalised |
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Ulk2 KO MEF (SV40) |
Mouse embryonic fibroblast, autophagy, Ulk2 -/- knockout SV40 immortalised |
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Ulk1 KO MEF (SIM) |
Mouse embryonic fibroblast, autophagy, Ulk1 -/-knockout spontaneously immortalised |
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Ulk1 KO MEF (SV40) |
Mouse embryonic fibroblast, autophagy, Ulk1 -/- knockout SV40 immortalised |
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Ulk/1/2 WT MEF (SIM) |
Mouse embryonic fibroblast, autophagy, Unc51-like kinase spontaneously immortalised |
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Ulk2 KO MEF (SIM) |
Mouse embryonic fibroblast, autophagy, Ulk2 -/- knockout spontaneously immortalised |
Key references
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McAlpine F, Williamson LE, Tooze SA, Chan EY. Regulation of nutrient-sensitive autophagy by uncoordinated 51-like kinases 1 and 2. Autophagy 2013 Mar 9(3):361-73 PMID:23291478
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Chan EY, Longatti A, McKnight NC, Tooze SA. Kinase-inactivated ULK proteins inhibit autophagy via their onserved C-terminal domains using an Atg13-independent mechanism. Mol Cell Biol. 2009 Jan;29(1):157-71. Epub 2008 Oct 20. PubMed PMID: 18936157; PubMed Central PMCID: PMC2612494.
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Chan EY, Kir S, Tooze SA. siRNA screening of the kinome identifies ULK1 as a multidomain modulator of autophagy. J Biol Chem. 2007 Aug 31;282(35):25464-74. Epub 2007 Jun 26. PubMed PMID: 17595159.
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Young AR, Chan EY, Hu XW, Köchl R, Crawshaw SG, High S, Hailey DW, Lippincott-Schwartz J, Tooze SA. Starvation and ULK1-dependent cycling of mammalian Atg9 between the TGN and endosomes. J Cell Sci. 2006 Sep 15;119(Pt 18):3888-900. Epub 2006 Aug 29. PubMed PMID: 16940348.