Figure 4. praja2 is required for tumour growth in vivo.

(a) U87MG cells transiently transfected with siRNAs were subcutaneously implanted into CD1 nude mice. Four weeks later, the mice were killed and the tumours excised and weighed. Tumour sections were fixed and doubly stained with hematoxylin/eosin or subjected to immunohistochemistry with anti-Ki67 antibody. Scale bar, 20 μm. (b) Cumulative data of tumour weight are expressed as mean±s.e.m. Twenty mice for each experimental group were used. *P<0.01 versus control siRNA, Student’s t-test. (c) Lysates from tumour lesions described in a were immunoblotted with anti-p21 (cyclin-dependent kinase (CDK) inhibitor) and anti-β actin antibodies. (d) Quantitative analysis of p21 levels in tumour lysates are expressed as mean±s.e.m. of three independent experiments. *P<0.01 versus control siRNA, Student’s t-test. (e) U87MG cells transiently transfected with control siRNA or praja2 siRNA were stereotaxically implanted into the brain (left caudate nucleus) of nude mice. Tissue sections from tumour lesions were stained with hematoxylin/eosin or immunostained with anti-Ki67. Scale bar, 400 μm. (f) Quantitative analysis of the experiments shown in e. Data are expressed a mean value±s.e.m. (Control siRNA: n=15, praja2 siRNA: n=10). *P<0.01 versus control siRNA, Student’s t-test. (g) FLT-PET analysis was performed at 2 and 4 weeks post-implantation. (h) Quantitative analysis of the experiment (at 2 weeks) shown in g. Data are expressed as mean value +/- s.e.m. *P<0.05 versus control siRNA, Student’s t-test.