Phosphorylation of 14-3-3ζ links YAP transcriptional activation to hypoxic glycolysis for tumorigenesis

a Subcellular fractionation analyses examining abundance of nuclear and cytoplasmic YAP protein expression in SW-1990 PDAC cells stimulated with hypoxia in the indicated times. GAPDH and Lamin B were used as internal control of cytoplasmic and nuclear extractions, respectively. SF serum-free, N normoxia, H hypoxia, WB western-blotting, NE nuclear extraction, CE cytoplasmic extraction, WCE whole cell extraction. Data are expressed as mean？±？s.d. of three independent experiments. *P？<？0.05. Two-sided Student’s t test was used to calculate the P value. b Representative immunfluorescence images of nuclear YAP localization in SW-1990 PDAC cells stimulated with hypoxia in the presence or absence of Flag-tagged wild-type 14-3-3ζ transfection for 6？h. Scale bar？=？25？μm. c Subcellular fractionation analyses detecting abundance of nuclear and cytoplasmic YAP protein expression in hypoxia-stimulated SW-1990 PDAC cells with or without Flag-tagged wild-type 14-3-3ζ transfection. d Coimmunoprecipitation assay evaluating the interaction between 14-3-3ζ and YAP in SW-1990 PDAC cells stimulated with hypoxia for 6？h. Data are expressed as mean？±？s.d. of three independent experiments. *P？<？0.05. Two-sided ANOVA with Bonferroni post hoc t test correction was used to calculate the P valu