Fig. 4. Targeting of phosphorylated L-plastin to areas of fast actin assembly. Transfected Vero cells expressing Ser5Glu (S/E), Ser5Ala (S/A) or wild-type L-plastin were processed for immunofluorescence double- or triple-staining. (Upper row) Cells expressing Ser5Glu variant were double-labelled with anti-cortactin and anti-sv-tag antibodies. Alexa-Fluor-488-coupled anti-mouse IgG (L-plastin S/E, green) and Texas-Red-coupled anti-rabbit IgG antibody (cortactin, red) served as secondary antibodies. The two right panels show merges of enlarged regions (boxes) of anti-sv-tag- and anti-cortactin-stained images. Bar, 15 µm. (Middle row) Cells expressing wild-type L-plastin were triple-labelled with Alexa-Fluor-350-phalloidin, anti-cortactin and anti-Ser5-P antibodies. Alexa-Fluor-488-coupled anti-mouse IgG (cortactin, green) and Texas-Red-coupled anti-rabbit IgG antibody (Ser5-P WT L-plastin, red) served as secondary antibodies. The right panel shows a merge of cortactin and anti-Ser5-P staining. Bar, 3 µm. (Lower row) Cells expressing L-plastin Ser5Ala were labelled with anti-cortactin and anti-sv-tag antibody. Secondary antibodies were used as described above (middle row). The right panel shows a merge of cortactin and sv-tag images. Bar, 3 µm.