A, TGLI1 protein was detected in patient GBMs sand direct xenografts. Resolution of TGLI1 and GLI1 proteins was enabled via prolonged 5.5% SDS-PAGE and WB with a GLI1 antibody. Stable GBM cells with TGLI1 and GLI1 were used to indicate the two GLI1 isoforms.
B, TGLI1-expressing GBM xenografts were significantly more aggressive in growth than the control and those with GLI1 (*,**, p<0.05). N=6 per group. Student t-test was conducted to determine p-values.
C, TGLI1-carrying GBM xenografts were highly vascularized. Three representative GBM xenografts are shown.
D, TGLI1-expressing xenografts contained significantly more blood vessels that those with GLI1. IHC was conducted to detect CD31 and derive microvessel density using a standard method [22, 25]. Student t-test was conducted to determine p-values. Right lower panels show high-resolution images.
E, TGLI1-expressing tumors were more proliferative than control and those with GLI1. IHC was conducted to detect Ki-67. Student t-test was used to determine p-values. Representative Ki-67-stained xenografts are shown in the right panels. Arrows point to brown Ki-67-positive tumor nuclei.
F, No significant difference in Akt activation status was observed among the three U87MG lines, as indicated by western blotting.