Phosphorylation of Ser-11 of AnxA2 interferes with S100A10 complex formation in MDCK cells. Subcellular localization of different AnxA2 mutants in MDCK cells stably transfected with the tetracycline-regulated XM-AnxA2 construct that induces AnxA2/S100A10 aggregates. MDCK-tTA-XM cells were cultivated either in (A) tetracycline-containing medium or in (B-E) the absence of tetracycline to induce expression of the XM construct. Subsequently, cells were transfected with the respective GFP-tagged AnxA2 constructs ([A-B] WT, [C] S11A, [D] S11D) or (E) a GFP expression vector as control. Twenty-four hours after transfection, cells were fixed, permeabilized, and stained with anti-S100A10 antibodies (clone H21) that recognize the XM derivative. WT- and S11A-AnxA2 but not S11D-AnxA2 are recruited to these clusters. Scale bars represent 10 µm. (F) Colocalization coefficients of S100A10-stained XM clusters and the respective AnxA2-GFP constructs of 7 single cells per experiment were determined using BioImageXD software, and statistical significance of 3 independent experiments was analyzed by 1-way ANOVA with Bonferroni test. Bars represent mean ± SEM.