Fig. 2

Downregulation of MYPT1 leads to Hippo pathway deactivation in ovarian cancer. a Gene Ontology term enrichment analyses of the genes whose expression levels correlated with the levels of MYPT1 in the GSE40595 and GSE38666 databases. Only biological process terms involving signaling pathways were selected. b Cumulative distribution of the Pearson’s correlation in the Hippo (left) or Wnt (right) pathway genes from GSE40595 (top) or GSE38666 (bottom). c Correlation of the expression levels of the NF2 and Hippo targets BIRC5, CTGF, GLI2 and FGF1 with the expression levels of MYPT1 in the GSE40595 and GSE38666 databases, in tumoral and non-tumoral tissue. Data were analyzed using Pearson’s R correlation. *, P < 0.05; **, P < 0.01; ***, P < 0.001. d Western blot showing the protein levels of MYPT1 in ES-2, SKOV3 and OVCAR8 cells expressing EV or shMYPT1. e Analysis of the MYPT1 expression level by RT-qPCR in ES-2, SKOV3 or OVCAR8 cells expressing EV or shMYPT1. f Heatmaps showing the z-scores of Hippo pathway gene expression obtained from TaqMan Array Human Hippo Signaling Pathway 96-well fast plates containing probes against Hippo pathway genes. Genes are sorted according to decreasing z-scores in the EV-expressing cells independently for each cell line. g Analysis of the NF2 expression level by RT-qPCR in ES2, SKOV3 or OVCAR8 cells. h Left, western blot showing the activation status of the Hippo signaling pathway in SKOV3 or OVCAR8 ovarian cancer cells expressing shMYPT1 or EV. Protein levels of pNF2 (S518), NF2, pMST1/2 (T180/183), pLATS1 (T1079), LATS, pYAP (S127), YAP, MYPT1 and α-tubulin are shown. Right, scheme showing the main components of the Hippo pathway and their activity with and without MYPT1. i Analysis of the expression of several Hippo pathway target genes, including BIRC5, CTGF, FGF1 and GLI2, by RT-qPCR in ES-2, SKOV3 or OVCAR8 ovarian cancer cells expressing shMYPT1, miR-30b or EV