Figure 2

Systematic overview and functional implications of nuclear YAP1/TAZ levels in tumor specimens of soft tissue and bone. (A,B) Immunohistochemical spectrum of tumor tissue specimens summarized as bar charts (intensity score) and (C,D) box plots (proportion score; shown are whiskers from minimum to maximum, 25th percentile, median and 75th percentile; +represents the mean of positive cells) for nuclear YAP1 and TAZ. Nuclear immunoreactivity was assessed using a semiquantitative score (0, negative; 1, weak; 2, moderate; and 3, strong) defining the staining intensity in the positive control (hepatocellular carcinoma) as strong. (E) Overall immunopositivity for nuclear YAP1 and TAZ. Only tumors with at least moderate staining (semiquantitative score ≥2) and ≥30% positive cells were considered positive for the purposes of the study. (F) Concordance of nuclear YAP1 and nuclear TAZ IHC-positivity. (G) Nuclear expression of YAP1 and TAZ in sarcoma cell lines. Immunoblotting demonstrates separated nuclear (N) and cytoplasmic (C) YAP1 and TAZ protein fractions in MLS1765-92 myxoid liposarcoma, CME-1 synovial sarcoma, ST88-14 malignant peripheral nerve sheath tumor and TC-32 Ewing sarcoma cells. Nuclear localization of YAP1/TAZ serves as indicator of YAP1- and TAZ-mediated transcriptional activity. Histone H3 and GAPDH were used as controls for the nuclear and cytoplasmic fraction, respectively. One of at least three independent experiments with comparable results is shown.