Fig. 7. NEK2 is a clinically relevant YAP/ MMB target in uveal melanoma.

A The significance for association with overall survival (OS) of uveal melanoma patients was calculated by stratifying for expression quintiles of YAP/MMB target genes. This analysis identified NEK2 as an MMB-target gene whose high expression is significantly associated with poor survival of uveal melanoma patients. See also Supplementary Table 6. B Kaplan–Meier survival analysis of uveal melanoma (UVM) and skin cutaneous melanoma patients (SKCM) in the TCGA datasets stratified by quintile NEK2 expression. C The expression of YAP and NEK2 after siRNA-mediated depletion of YAP/TAZ or expression of MY-COMP in 92.1 cells was analyzed by RT-qPCR. N = 3 replicates. Error bars indicate SD. Student’s t test. D 3C-qPCR chromatin capture experiments were performed to assess the interaction between the YAP bound enhancer and the NEK2 promoter. The top shows the CUT&RUN tracks of YAP, H3K4me1, K3K4me3, H4Ac and H3K27Ac at the NEK2 locus demonstrating binding of YAP to an active enhancer 60 kb telomeric of the NEK2 promoter. The arrows indicate the location of primers used in the 3C-qPCR analysis. The constant anchor primer (a) is indicated in green. Interaction frequencies were determined by qPCR after HindIII digestion and re-ligation relative to the ligation products from the BAC DNA covering the same genomic region. Data shown are from one representative experiment from two independent experiments.