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Figure 3 | BMC Cancer

Figure 3

From: The role of versican G3 domain in regulating breast cancer cell motility including effects on osteoblast cell growth and differentiation in vitro– evaluation towards understanding breast cancer cell bone metastasis

Figure 3

Expression of versican G3 domain regulated MC3T3-E1 cells growth and differentiation. (a) Vector- and G3- transfected MC3T3 cells (2 × 104) were inoculated in 6-well culture dishes containing 10% FBS/AMEM and cultured for 12 h. After cell attachment, the cells were cultured with or without 1 ng/ml TGF-β1 for 5 d. Cells were also counted by light microscope in 1, 3, 5 days. n = 6, * p<0.05, ** p<0.01, analyzed with t-test. (b) The G3- and vector-transfected MC3T3-E1 cells (1 × 103) were inoculated in 96-well culture dishes and cultured in AMDM medium containing 1 ng/ml TGF-β1 or not for 7 days. Proliferation assays performed with WST-1 assay. All groups compared with vector control cells, n = 8, * p<0.05, ** p<0.01, analyzed with t-test. (c) The vector- and G3-transfected MC3T3-E1 cells were seeded at 8 x104 cells/well in a 6 well plate. Cells were maintained in 10% FBS/AMEM medium containing 1 ng/ml TGF-β1 for 21 days. The medium was changed every 3 days. Cells were lysised and processed to ALP ELISA Assay. All groups compared with vector control cells, n = 6, * p<0.05, ** p<0.01, analyzed with t-test. (d) Typical pictures showed ALP staining of vector- and G3-transfected MC3T3-E1 cells maintained in 10% FBS/AMEM medium containing 1 ng/ml TGF-β1 for 21 days. (e) The vector- and G3-transfected MC3T3-E1 cells were maintained in 10% FBS/AMEM medium with or without 1 ng/ml TGF-β1 for 3 days. All cells were lysed and subjected to immunoblotting with antibodies to pEGFR, pAKT, pSAPK/JNK, GSK-3β (S9P) and β-actin.

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