The protein 4.1 (P4.1R) (Gascard et al., 1998; Calinisan et al., 2006) is a multifunctional protein that localizes to the membrane skeleton and stabilizes cell shape and membrane mechanical properties, such as deformability and stability, via lateral interactions with other proteins of the membrane cytoskeleton and with the membrane itself (with its N-terminal FERM domain). Preliminary observations demonstrated that HEK cells transiently trasfected with P4.1 R show an increased number of filopodia. To better evaluate this phenomenon we established a protocol of Correlative Light-Scanning Electron Microscopy. We identified cells grown on a etched cover slip and transfected with a P4.1-IRES-GFP vector, by live scanning confocal microscopy and then analysed the same cells at the ultrastructural level to precisely evaluate the density, distribution and features of the filopodia. ICln is a protein known to interact with the FERM domain of P4.1 (Tang and Tang, 1998). We co-transfected the cells with P4.1-IRES-GFP and with a ICln-IRES-DsRed vector and observed a milder phenotype. We then hypothesized that this interaction might interfere with the activation of the pathways leading to filopodia emission.

Correlative light-scanning electron microscopy on P4.1 overexpressing hek cells / S. Rodighiero, E. Mascia, D. Marchesi, C. Bazzini, G. Bongiorno, M. Francolini. ((Intervento presentato al 2. convegno Transalp'nano 2010 tenutosi a Como nel 2010.

Correlative light-scanning electron microscopy on P4.1 overexpressing hek cells

C. Bazzini;M. Francolini
Ultimo
2010

Abstract

The protein 4.1 (P4.1R) (Gascard et al., 1998; Calinisan et al., 2006) is a multifunctional protein that localizes to the membrane skeleton and stabilizes cell shape and membrane mechanical properties, such as deformability and stability, via lateral interactions with other proteins of the membrane cytoskeleton and with the membrane itself (with its N-terminal FERM domain). Preliminary observations demonstrated that HEK cells transiently trasfected with P4.1 R show an increased number of filopodia. To better evaluate this phenomenon we established a protocol of Correlative Light-Scanning Electron Microscopy. We identified cells grown on a etched cover slip and transfected with a P4.1-IRES-GFP vector, by live scanning confocal microscopy and then analysed the same cells at the ultrastructural level to precisely evaluate the density, distribution and features of the filopodia. ICln is a protein known to interact with the FERM domain of P4.1 (Tang and Tang, 1998). We co-transfected the cells with P4.1-IRES-GFP and with a ICln-IRES-DsRed vector and observed a milder phenotype. We then hypothesized that this interaction might interfere with the activation of the pathways leading to filopodia emission.
2010
Settore BIO/14 - Farmacologia
Universita' degli Studi di Milano
Fondazione Filarete
Universita' degli Studi di Milano Bicocca
Politecnico di Milano
Provincia di Varese
NanoScience Rhone-Alpes
Correlative light-scanning electron microscopy on P4.1 overexpressing hek cells / S. Rodighiero, E. Mascia, D. Marchesi, C. Bazzini, G. Bongiorno, M. Francolini. ((Intervento presentato al 2. convegno Transalp'nano 2010 tenutosi a Como nel 2010.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/162051
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