Figure 6

Somatic hSH3BGRL mutant R76C resembled to mSH3BGRL promotes lung metastasis. (a) Lysates from CHO cells stably transfected with hSH3BGRL or its somatic mutant R76C and immunoblotted with the indicated antibodies, respectively. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as a loading control. The indicated protein relative expression level is quantified under the immediate panel. (b) In total, 1x10⁶ CHO hSH3BGRL- or its hSH3BGRL-R76C-overexpressing cells (R76C) were injected intravenously into the tail vein of nude mice. After 17 days, mice were killed and their lungs were photographed and scored for metastatic tumor nodules; mean±s.d., n=5, ***P<0.0001, unpaired Student's t-test. (c) Lysates from MDA-MB-231 cells stably transfected with empty vector (Vector), wild-type hSH3BGRL and its somatic mutant R76C and immunoblotted with the indicated antibodies. The averaged relative protein expression level is quantified and shown under the immediate panels. (d) Effect of hSH3BGRL and its somatic mutation R76C to lung metastasis in MDA-MB-231 cells. In total, 1x10⁶ stably transfected cells in (c) were injected into nude mice through tail vein. After 15 days injection, the assayed mice were killed and the dissected lungs were photographed and the metastatic nodules were counted. mean±s.d., n=4, **P<0.001; ***P<0.0001, unpaired Student's t-test.