WW domains are dispensable to YAP1 phosphorylation catalyzed by LATS1/2. (A) The representative western blotting shows the Ser-127 phosphorylation catalyzed by the MST2−LATS2 kinase cascade on YAP1 and the W4A mutant in HEK293T cells. The inserted numbers show the relative Ser-127 phosphorylation levels normalized by the YAP1 content. (B) The representative western blotting shows the Ser-127 phosphorylation catalyzed by the MST2−LATS1 kinase cascade on YAP1, the W4A mutant, and the ΔWWs mutant in HEK293T cells. The inserted numbers show the relative Ser-127 phosphorylation levels normalized by the YAP1 content. (C) The co-IP experiment shows the interaction of MYC-tagged TEAD1 with HA-tagged YAP1 or the ΔWWs mutant in HEK293T cells. TCL, total cell lysates. (D) The representative western blotting shows the depletion of YAP1 expression in YAP1-knockout (KO) HeLa cells. (E) Representative images show subcellular distributions of the full-length YAP1, the shorter amino terminus that contains TBD, and the extended amino terminus that contains TBD and the tandem WW domains (TBD−WWs) in HEK293T cells (the yellow line indicates the nuclear outline). The scatter chart shows their nucleocytoplasmic ratios (representative of three independent experiments, n represents the total number of the measured cells, two-tailed ANOVA test). (F) Representative images show subcellular distributions of the cNLS fused YAP1 and the ΔWWs mutant in HEK293T cells. The cNLS is derived from the SV40 virus. TAD, transcriptional activity domain. Source data are available for this figure: SourceData FS1.