Actin Cytoskeleton and Adhesive Structures

Cells were treated as described in (Majewski et al. 2011 (link)), and then stained with Alexa 488-conjugated phalloidin to visualize the actin cytoskeleton. Staining for adhesive structure markers was also performed using monoclonal antibody against vinculin (Sigma-Aldrich, USA) and polyclonal antibody against talin (Santa Cruz, USA) at a dilution of 1:40. For negative controls, primary antibody was omitted. Images were collected with the Leica TCS SP5 confocal laser scanning microscope equipped with a 63× HCX oil CS UV 1.4 oil-immersion objective.

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Pavlyk I., Rzhepetskyy Y., Jagielski A.K., Drozak J., Wasik A., Pereverzieva G., Olchowik M., Kunz-Schugart L.A., Stasyk O, & Redowicz M.J. (2014). Arginine deprivation affects glioblastoma cell adhesion, invasiveness and actin cytoskeleton organization by impairment of β-actin arginylation. Amino Acids, 47(1), 199-212.

Publication 2014

monoclonal antibody against vinculin TCS SP5 confocal laser scanning microscope

Actin cytoskeleton Antibody Cells Confocal laser scanning microscope Dilution Immersion Monoclonal antibody Phalloidin Talin Vinculin

Corresponding Organization :

Other organizations : Instytut Biologii Doświadczalnej im. Marcelego Nenckiego, University of Warsaw, Polish Academy of Sciences

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Variable analysis

independent variables

Treatment of cells as described in (Majewski et al. 2011)

dependent variables

Visualization of the actin cytoskeleton using Alexa 488-conjugated phalloidin staining
Staining for adhesive structure markers using monoclonal antibody against vinculin and polyclonal antibody against talin

control variables

Dilution of primary antibodies (1:40)

controls

Negative controls: Primary antibody was omitted

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