Fig. 3. Ethanol reorganizes focal adhesions and decreases their levels. Astrocytes were exposed to 100 mM ethanol for an hour. (A) Actin (a,c,e) and paxillin (b,d,f) were detected by immunofluorescence. Control astrocytes show a punctuate pattern of paxillin (b). Ethanol treatment causes a paxillin reorganization at the cell periphery (d) and dramatically decreases its content when cells are densely packed (f). (B) A representative western blot of paxillin from control- and ethanol-treated cells, at different times of alcohol exposure. (C) Densitometric quantification of paxillin staining at two ethanol concentrations (100 and 200 mM for 24 hours). Equal amount of protein was used (50 µg/well). The values represent means ±s.d. from four different cultures and they are expressed as percentages of the control value. Asterisks indicate significant differences with control sample (**P<0.001). (D) Astrocytes were double-stained for F-actin (a,c,e) and phosphotyrosine (b,d,f). Ethanol-exposed cells show a cortical reorganization (d) and a reduction in phosphotyrosine staining of the focal adhesion proteins (f). Bars, 10 µm.