Invadopodia biogenesis is regulated by caveolin-mediated modulation of membrane cholesterol levels
Article first published online: 31 OCT 2008
© 2008 The Authors Journal compilation © 2009 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd
Journal of Cellular and Molecular Medicine
Volume 13, Issue 8b, pages 1728–1740, August 2009
How to Cite
Caldieri, G., Giacchetti, G., Beznoussenko, G., Attanasio, F., Ayala, I. and Buccione, R. (2009), Invadopodia biogenesis is regulated by caveolin-mediated modulation of membrane cholesterol levels. Journal of Cellular and Molecular Medicine, 13: 1728–1740. doi: 10.1111/j.1582-4934.2008.00568.x
- Issue published online: 26 NOV 2009
- Article first published online: 31 OCT 2008
- Received: July 21, 2008; Accepted: October 24, 2008
Fig. S1. Effect of cholesterol depletion on pre-formed invadopodia. A375MM cells were plated on gelatin and left untreated (NT) or incubated with 4 μM lovastatin/0, 25 mM mevalonate (L+M) for 16 hrs. Treated cells were then subjected or not to acute cholesterol extraction with 10 mM methyl-betacyclodextrin (MCD) for 40 min., incubated for further 3 hrs and then fixed and stained with phalloidin. The percentage of cells with invadopodia over total cells was then calculated, considering at least 30 random fields per each sample.
Fig. S2. Phospho-Src levels in caveolin 1Y14D- and caveolin 1Y14F-transfected cells. A375MM caveolin 1 knock-down cells were transfected with GFP-tagged caveolin 1Y14D and caveolin 1Y14F and plated on unconjugated gelatin coated coverslips for 16 hrs. Samples were then labelled with an antibody recognizing phosphorylated Src and Alexa Fluor 633-conjugated phalloidin. Levels of phosphorylated Src were calculated as the ratio fluorescence/cell area, considering transfected and untransfected (NT) cells in the same field and expressed as a percentage of pSrc intensity in untransfected cells. The standard deviation was calculated among 20 fields per each sample.
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