title: Measuring Expression Levels of Endogenous Gli Genes by Immunoblotting and Real-Time PCR.
abstract: Gli proteins are transcription factors that mediate the transcriptional effects of Hedgehog signaling in vertebrates. The activities of Gli2 and Gli3 are regulated primarily by posttranslational modifications, while Gli1 is mostly regulated at the transcriptional level. Detection of endogenous Gli proteins had been hampered by lack of good antibodies, but this problem has been mostly resolved in recent years. In this chapter we describe methods of detecting expression of endogenous Gli genes in whole-cell lysates and in subcellular fractions and also provide protocols for the measurement of Gli mRNA levels by quantitative real-time reverse transcriptase PCR (qPCR).
authors.affiliation: Department of Cell Biology, Nencki Institute of Experimental Biology, 02-093, Warszawa, Poland, p.niewiadomski@nencki.gov.pl.
authors.forename: Pawel
authors.initials: P
authors.lastname: Niewiadomski
authors.forename: Rajat
authors.initials: R
authors.lastname: Rohatgi
mesh_terms: D000818:Animals; D015870:Gene Expression; D015151:Immunoblotting; D042541:Intracellular Space; D051379:Mice; D041681:NIH 3T3 Cells; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism*; D010766:Phosphorylation; D021381:Protein Transport; D060888:Real-Time Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*; D000071676:Zinc Finger Protein GLI1
chemical_list: D015513:Oncogene Proteins; D015534:Trans-Activators; D000071676:Zinc Finger Protein GLI1