As previously reported [46], deletion of Pals1 in the colorectal cancer cell line HCT116 results in disturbed tight junction formation as evaluated by staining for ZO-1 (Fig. 1A) and Occludin (Supplementary Fig. 1A), a loss of its canonical interaction partners PATJ and Crb3a from cell–cell contacts (Supplementary Fig. 1A, B), enhanced collective cell migration (Supplementary Fig. 1C) and increased invasion through extracellular matrix (Supplementary Fig. 1D). Here, TJP1 is linked to colorectal cancer.