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Cortical plaques

Cortical plaques. Lambotin et al , J Cell Science 2005. Ezrin. Actine (nid d’abeille). GTPases. Doulet et al , J Cell Biol 2006. PI3K. Nécessaire à l’adhésion "forte" en condition de flux (G. Mikaty, G. Dumenil & coll). Lambotin et al , J Cell Science 2005. β -arrestins. A. B.

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Cortical plaques

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  1. Cortical plaques Lambotin et al, J Cell Science 2005 Ezrin Actine (nid d’abeille) GTPases Doulet et al, J Cell Biol 2006 PI3K Nécessaire à l’adhésion "forte" en condition de flux (G. Mikaty, G. Dumenil & coll) Lambotin et al, J Cell Science 2005

  2. β-arrestins A B C • MapK • Src • Akt Lan Ma and Gang Pei, Journal of Cell Science 120, 213-218 (2007)

  3. β-arrestins - SiRNA Beta arrestine 2 GFP ezrin βarrestin 1/2 siRNA Ezrin recruitment Expression is reduced by Control siRNA 100% 80% βarrestin 1/2 2,5 fold 60% Ezrin Actin 40% 20% 0% Control Arrestin 1+2 FLNA sirRNA GRK2 sirRNA siRNA sirRNA

  4. β-arrestins - SiRNA Beta arrestine 2 GFP ezrin βarrestin 1/2 siRNA Ezrin recruitment Expression is reduced by Filamin A siRNA Control siRNA GRK2 siRNA 100% 80% βarrestin 1/2 2,5 fold 60% Ezrin Filamin A 7,5 fold 40% GRK2 20% 3 fold 0% Actin Control Arrestin 1+2 FLNA sirRNA GRK2 sirRNA siRNA sirRNA

  5. Ezrin recruitment 100 80 60 % of recruiting colony 40 20 0 Control Phentolamine 10 uM Propranolol 10 uM Recepteur adrénergique ezrin Beta2 AR - YFP α-antagonist β-antagonist

  6. Ezrin recruitment 100 80 60 % of recruiting colony 40 20 0 Ct 0,0001 0,01 1 10 Isoproterenol (μM) β-agonist Recepteur adrénergique ezrin Beta2 AR - YFP +

  7. + Ezrin recruitment 100 80 60 % of recruiting colony 40 20 0 Ct 0,0001 0,01 1 10 Isoproterenol (μM) β-agonist Recepteur adrénergique ezrin Beta2 AR - YFP

  8. Ezrin recruitment Ezrin recruitment 100% 100% 80% 80% 60% 60% 40% 40% 20% 20% 0% 0% IL8 MIP Isoproterenol Propranolol • 10µM 10µM 1µM 1µM - - + - + AT2 SDF1 control Isoproterenol 1uM Isoproterenol 100nM 5 à10 x Kd Recepteur adrénergique

  9. Ezrin recruitment 100 80 60 % of recruiting colony 40 20 0 PKA-PS Forkolin 10-5 M PKA-PS Forkolin 10-6 M Control 20uM 10uM Signaling des protéines G

  10. Activation partielle

  11. Billes coatées, cellules hCMEC/D3 contraste actin ezrin Isoproterenol β1 agoniste Phenylephrine α1 agoniste BSA ou Metoprolol

  12. Recrutment d’Ezrin Recrutment d’Ezrin 100 100 80 80 60 60 % recruiting colonies % recruiting colonies 40 40 20 20 0 0 Ct 0,0001 0,01 1 10 Ct 0,0001 0,01 1 100 Iso + Isoproterenol (μM) Phenylephrine (μM) Phe 10:10 5 4 3 Ezrin phosphorylation fold increase 2 1 0 Isoproterenol Ct Effet des agonistes adrénergiques Isoproterenol Ct Nm - + - + P-Ezrin P-Moesin

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