Figure 6: Enforced TAZ expression promotes GBM-associated malignant phenotypes.

A: GBM1B and A172 cells were infected with lentiviruses harboring TAZ (FLAG-tagged) cDNA or no cDNA insert as the control to established stable cell lines. Transgene expression was measured by western blotting.

B and C: Validated TAZ-Up genes were analyzed by qRT-PCR (B) in GBM1B and GBM1B-TAZ cells, and by western blotting (C) using whole cell lysates from GBM1B and A172 cells with +/− enforced TAZ expression.

D: Cell growth curves were drawn from the number of trypan blue stained GBM1B and A172 cells with +/− enforced TAZ expression.

E: Colony formation was quantified in GBM1B and GBM1B-TAZ cells in medium with +/− EGF for 10 days.

F: GBM1B with +/− enforced TAZ expression were grown in medium without growth factors for 48 hours, and EGF release in medium was measured by ELISA.

G: GBM1B cells were grown in soft agar with EGF-containing medium or EGF-free conditioned medium (24h) from GBM1B and GBM1B-TAZ cells. Colony formation after 10 days was quantified.

H: GBM1B and GBM1B-TAZ cells were plated onto laminin-coated Transwell membranes. Cell migration was compared after 24 h by quantifying DAPI+ cells per field.

I: A172 and A172-TAZ cells were subjected to scratch wound healing assay to quantify migrating cells in the scratch area as shown in Figure S7.

J: GBM1B and GBM1B-TAZ cells received +/− irradiation (1 or 3 daily doses of 3 Gy). Clonogenic survival was quantified and normalized to untreated control cells.

K: 10,000 viable A172 and A172-TAZ cells were transplanted into mouse brains (n=5). Coronal brain sections (20 μm, with H&E staining) were shown (post-implantation day 76, Bar = 0.5 mm (left) and 50 μm (right); arrowhead: necrotic area).

L and M: 10,000 viable GBM1B and GBM1B-TAZ cells were transplanted into mouse brains (n=5). Coronal brain sections (20 μm, with H&E staining) were shown (day 60 post implantation, Bar = 0.5 mm) with tumor size quantification (L). hNu immunostaining (M) was used to detect and quantify tumor cells invading the contralateral corpus callosum area (marked by black rectangles in L). hNu+ cell numbers were quantified in 5 fields with equal sizes as marked underneath the picture (Bar = 100 mm).

Data are represented as mean ± SEM (*: p < 0.01).