Figure 1: EGF induces TAZ expression in GBM cells.

A: TAZ (WWTR1) mRNA levels positively correlate with EGF and FGF2 levels in GBM specimens from the TCGA database (n=454).

B: GBM1B cells were depleted of growth factors for 16 hours and treated with RTK ligands as indicated for 1h or 5min for testing TAZ induction or phosphorylation of their receptors, respectively (Con: RTK ligand-untreated cells).

C: GBM1B cells were deprived from growth factors for 16 hours and treated with EGF for 2 hours. TAZ mRNA was measured by qRT-PCR.

D: After growth factor depletion for 16 hours, GBM cells as marked received EGF treatment for the indicated times (Con: untreated cells). TAZ protein was quantified by western blotting.

E and F: After growth factor depletion for 16 hours, GBM1B cells were treated with EGF for the indicated times (Con: untreated cells). TAZ western blotting was performed using proteins from nuclear and cytosol fractions (E). Cells with +/− EGF treatment (4 hours) were subjected to TAZ immunostaining and quantification of TAZ signal intensity in DAPI+ nuclei (F; DAPI: nuclear counterstaining; Bar = 10 μM; n=50)

G and H: The same cells as used in E were subjected to TAZ ChIP-PCR with IgG as the control (G, NC: negative control regions randomly selected from the genome). CTGF and MYC transcription was also quantified (H).

Protein fold expression normalized to β-Actin, total levels of receptors or TBP (E) is shown below each lane. Data are represented as mean ± SEM (*: p < 0.01).