Analyzing RB and E2F during the G1-S transition

Methods Mol Biol. 2014:1170:449-61. doi: 10.1007/978-1-4939-0888-2_24.

Abstract

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.

MeSH terms

  • Animals
  • E2F Transcription Factors / metabolism*
  • Equipment Design
  • G1 Phase*
  • Humans
  • Immunoblotting / methods
  • Nucleic Acid Hybridization / methods
  • Phosphorylation
  • RNA / genetics
  • RNA / isolation & purification
  • Retinoblastoma Protein / metabolism*
  • Transcriptional Activation*

Substances

  • E2F Transcription Factors
  • Retinoblastoma Protein
  • RNA