Podosome reformation in macrophages: assays and analysis
Standard
Podosome reformation in macrophages: assays and analysis. / Cervero, Pasquale; Panzer, Linda; Linder, Stefan.
in: Methods Mol Biol, Jahrgang 1046, 01.01.2013, S. 97-121.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Podosome reformation in macrophages: assays and analysis
AU - Cervero, Pasquale
AU - Panzer, Linda
AU - Linder, Stefan
PY - 2013/1/1
Y1 - 2013/1/1
N2 - Podosomes are multifunctional organelles of invasive cells that combine several key abilities including cell-matrix adhesion, extracellular matrix degradation, and mechanosensing. In combination with their high turnover rates that allow quick adaptation to the pericellular environment, podosomes are likely to play important roles during invasive migration of cells. Primary human macrophages constitutively form numerous podosomes and are thus an ideal system for the quantitative study of podosome dynamics. This protocol describes assays for the study of podosome dynamics, namely, reformation of podosomes, in fixed and living cells, with subsequent software-based analyses allowing the extraction of quantitative parameters such as the number of podosomes per cell, podosome density, and half times for podosome disruption and reformation. Moreover, we describe the preparation of podosome-enriched cell fractions and their analysis by immunoblotting.
AB - Podosomes are multifunctional organelles of invasive cells that combine several key abilities including cell-matrix adhesion, extracellular matrix degradation, and mechanosensing. In combination with their high turnover rates that allow quick adaptation to the pericellular environment, podosomes are likely to play important roles during invasive migration of cells. Primary human macrophages constitutively form numerous podosomes and are thus an ideal system for the quantitative study of podosome dynamics. This protocol describes assays for the study of podosome dynamics, namely, reformation of podosomes, in fixed and living cells, with subsequent software-based analyses allowing the extraction of quantitative parameters such as the number of podosomes per cell, podosome density, and half times for podosome disruption and reformation. Moreover, we describe the preparation of podosome-enriched cell fractions and their analysis by immunoblotting.
KW - Biological Assay
KW - Cell Fractionation
KW - Cell Movement
KW - Cell-Matrix Junctions
KW - Extracellular Matrix
KW - Humans
KW - Macrophages
KW - Molecular Biology
KW - Neoplasm Invasiveness
U2 - 10.1007/978-1-62703-538-5_6
DO - 10.1007/978-1-62703-538-5_6
M3 - SCORING: Journal article
C2 - 23868584
VL - 1046
SP - 97
EP - 121
JO - Methods Mol Biol
JF - Methods Mol Biol
SN - 1064-3745
ER -