ABSTRACT
Nuclear microenvironments are architecturally organized subnuclear sites where the regulatory machinery for gene expression, replication, and repair resides. This compartmentalization is necessary to attain required stoichiometry for organization and assembly of regulatory complexes for combinatorial control. Combined and methodical application of molecular, cellular, biochemical, and in vivo genetic approaches is required to fully understand complexities of biological control. Here we provide methodologies to characterize nuclear organization of regulatory machinery by in situ immunofluorescence microscopy.
Subject(s)
Intracellular Space/metabolism , Intranuclear Space/metabolism , Transcription Factors/metabolism , Animals , Cell Adhesion , Cell Culture Techniques , Computational Biology , Fluorescence Recovery After Photobleaching , Intermediate Filaments/metabolism , Metaphase , Microscopy , Nuclear Matrix/metabolism , Protein TransportABSTRACT
Regulatory machinery for gene expression, replication, and repair are architecturally organized in nuclear microenvironments. This compartmentalization provides threshold concentrations of macromolecules for the organization and assembly of regulatory complexes for combinatorial control. A mechanistic under standing of biological control requires the combined application of molecular, cellular, biochemical, and in vivo genetic approaches. This chapter provides methodologies to characterize nuclear organization of regulatory machinery by in situ immunofluorescence microscopy.
