p190A interacts with cortactin via the PLS domain. (A) Huh7 cells transfected with both mcherry-cortactin and indicated HA-tagged p190A plasmids were used to perform immunoprecipitation (IP) with anti-HA beads. Presence of cortactin is analyzed by immunoblot using an anti-cortactin antibody. (B) Coimmunoprecipitation was performed on Huh7 lysates with IgG control (Ctl), anti-p190A, and anti-cortactin antibodies. PLA was performed on Huh7 cells transfected (C) or not (E) with the indicated constructs. Cells were stained for nuclei (Hoechst, blue). The green staining represents, as shown on each panel, the HA staining, GFP tag, or F-actin staining. The dots in the PLA panel represent locations where interaction between indicated proteins (stained using primary antibodies from mouse [m] or rabbit [rb] species) occurs; dots density is quantified in D. Values are expressed as the mean ± SEM of three independent experiments (n = 9 cells per condition). P-value from the ANOVA test is indicated. **, P < 0.01.