Analyzing RB and E2F During the G1–S Transition

  • Michael J. Thwaites
  • Matthew J. Cecchini
  • Frederick A. Dick
Part of the Methods in Molecular Biology book series (MIMB, volume 1170)


The G1/S-phase restriction point is an important landmark in the mammalian cell division cycle. The key regulator of the G1/S transition is the retinoblastoma gene product (pRB). It prevents the transcription of genes required for S-phase progression by repressing E2F transcription factors. An increase in Cdk phosphorylation of pRB causes the release of E2F transcription factors and advancement into S phase. Here we describe two simple techniques used to assess pRB phosphorylation and E2F transcription during G1/S progression.

Key words

Retinoblastoma Transcription Phosphorylation Gene expression E2F 


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Michael J. Thwaites
    • 1
    • 3
  • Matthew J. Cecchini
    • 1
    • 3
  • Frederick A. Dick
    • 1
    • 2
    • 3
    • 4
  1. 1.London Regional Cancer ProgramLondonCanada
  2. 2.Children’s Health Research InstituteWestern UniversityLondonCanada
  3. 3.Department of BiochemistryWestern UniversityLondonCanada
  4. 4.Cancer Research LabsLondonCanada

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