Fig. 6: GAB2 is regulated by PP1.

a Dephosphorylation detected by Western Blot. GAB2-pS210 signal relative to GAB2 is quantified on the right. Shown is one representative experiment out of three independent repeats (CalA: Calyculin A). Four independent repeats have been quantified on the right. p-Values are derived from a t-test (two-sided, paired). b Overexpressed GFP-14-3-3β is less associated with endogenous GAB2 upon PP1c treatment. Inhibition of PP1c with calyculin A (CalA) abolishes the effect. Quantification depicts results of three independent experiments with p-values obtained from t-tests (two-tailed, paired). For a, b: Samples were derived from the same experiment and gels/blots were processed in parallel. c Confocal microscopy images of pmKate2-Gab2 signal in live HeLa Kyoto cells are shown after the indicated treatments. Stimulus was added after 45 s, scale bar represents 25 µm. (see also Supplementary Movies 1-3) d Scheme of image analysis for calculating GAB2 enrichment at the plasma membrane. e Fold-enrichment of GAB2 at the plasma membrane, as determined by comparing 0 min and 5 min frames. Each point represents a single cell (n = 44/56/31 for EGF/PDP-Nal/PDPm-Nal). Statistics represent adjusted p-values of a two-sided Wilcoxon test. Boxplots highlight median and 1st/3rd quartile. Whiskers refer to the lowest and highest data points that are within a 1.5-fold interquartile range. Source data of Fig. 6a, b, and e are provided as a Source Data file.