PP2A_STRIPAK regulates Slik localization by phosphorylation to control moesin activation. (A) S2 cells stably expressing Slik^WT-GFP (white top row or green bottom row) upon the indicated treatment and stained for F-actin (red) and DNA (blue). (B) The ratio of Slik^WT-GFP intensity (cortex/cytoplasm) was quantified in a minimum of 40 cells/experiment in three independent experiments (see Fig. S3 D). Data are represented as mean ± SD (each dot represents the value for a single cell), one-way ANOVA. (C) Illustration of Slik (short isoform of 1,300 amino acids) and its kinase domain (KD), NCD, and CTD containing coiled-coil motifs (cc). Previously identified phosphosites (light blue; Panneton et al., 2015) and additional phosphosites (dark blue) are shown. (D) Cells expressing the indicated Slik-GFP constructs (white top row). The signal of the indicated Slik-GFP construct was pseudocolored in a rainbow heatmap to underline variations in its localization (bottom row). (E and F) The ratio of Slik-GFP intensity (cortex/cytoplasm) was quantified in S2 cells stably expressing the indicated constructs. 30 (E) or 40 (F) cells/condition in three (E) or two (F) independent experiments (see Fig. S3). Data are represented as mean ± SD (each dot represents the value for a single cell), two-tailed unpaired t test (E) or one-way ANOVA (F). (G) Western blot of S2 cells stably expressing the indicated Slik-GFP constructs and treated as indicated. (H) Relative p-moesin levels (normalized to moesin) were quantified in two independent experiments (circles). Data are represented as mean ± SD normalized on nontarget (NT) dsRNA. P values are as follows: NS, P > 0.05; *, P < 0.05; **, P < 0.01; ****, P < 0.0001. Scale bars, 5 µm. OE, overexpressed.