Figure 7.

Interaction of native CAP1 with cofilin isoforms in cells: (a) western blot analysis demonstrates the specificity of the primary isoform-specific anti-CFL1, anti-CFL2, and anti-CAP1 antibodies used in immunofluorescence proximity ligation assay (PLA) experiments. WCL: whole-cell lysate. Uncropped versions of the blots are shown in Supplementary Figures S4 and S5. (b) Duolink in situ PLA assay performed on Hs 578T cells, as described in the Section 4.8. Cells stained using a single primary antibody (CFL1 only, CFL2 only, and CAP1 only) are shown as negative controls. PLA signal (magenta) using pairs of CFL1/CAP1 and CFL2/CAP1 antibodies represents CAP1/cofilin interaction events. Cells were counter-stained with fluorescein isothiocyanate (FITC)–phalloidin for F-actin (green) and nuclear 4′,6-diamidino-2-phenylindole (DAPI, blue). Scale bar is 20 μm.