Extended Data Fig. 9: FAT1 loss results in dysregulation of YAP1 nuclear localization.
a,b Transient siRNA depletion of FAT1 increases the relative YAP1 nuclear localization ratio in RPE1-FUCCI cells (a; PTEMF fixation) and T2P cells (b (left panel); PFA fixation). Scale bars =10 µm. Representative images are shown. The right panel in b shows quantification of the results in T2P cells. Red bar =mean, 2-sided Mann-Whitney test. c, Transient siRNA depletion of FAT1 and LATS1 does not consistently alter the phosphorylation level of YAP1 in RPE1 and T2P cells. d, Plot showing the incidences of amplification and deep deletions of Hippo pathway members in the TRACERx 421 LUSC cohort. The effectors TEAD4 and WWTR1/TAZ are amplified in >10% and >35% of cases, respectively. e, The localization of the HA-epitope tag does not affect the ability to repress TEAD reporter transcriptional activity. Left, Scheme showing the location of the HA-epitope tag in different FAT1 expression constructs. The HA-epitope tag was located either at the N terminus or internally within the FAT1 construct. Right, Histogram showing normalized TEAD transcriptional activity upon overexpression of different FAT1 construct. The edge of histograms denotes the mean values, One-way ANOVA with Holm-Sidak multiple comparisons test, for each condition >95 cells were scored across 3 biological repeats.
