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Figure 5 | BMC Cancer

Figure 5

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 5

Expression of versican G3 domain regulated MC3T3-E1 cells apoptosis via EGFR/JNK pathway. (a) G3- and vector-transfected MC3T3-E1 cells (2 × 105) were inoculated in 6 well culture dishes with 10% FBS/AMEM medium. After cultured for 12 hours, all samples were treated serum free AMEM medium with or without 2 ng/ml TNF-α for 4 days. The survival cells were counted under light microscope and compared with the seeded cells to present the viability (%). All groups compared with vector control cells, n = 6, * p<0.05, ** p<0.01, analyzed with t-test. (b) G3- and vector-transfected MC3T3-E1 cells (1 x 103) were inoculated in 96-well culture dishes and cultured in 10%FBS/AMEM medium for 12 h. All samples were treated serum free AMEM medium with or without 2 ng/ml TNF-α for 7 days. All groups compared with vector control cells, n = 9, * p<0.05, ** p<0.01, analyzed with t-test. The cell viability was assayed by WST-1 assays. (c) G3- and vector-transfected MC3T3-E1 cells were treated with serum free AMEM medium with 2 ng/ml TNF-α for 24 hours and processed to Annexin V assays. (d) G3- and vector-transfected MC3T3-E1 cells were treated with serum free AMEM medium with 2 ng/ml TNF-α for 24 hours. All samples were lysed and subjected to immunoblotting with antibodies to pEGFR, pSAPK/JNK, GSK-3β (S9P) and β-actin. (e) G3- and vector-transfected MC3T3-E1 cells were treated with serum free AMEM medium with 2 ng/ml TNF-α, with or without 100 nM SP60025 for 4 days. Cell viability was assessed by WST-1 assays. The cell viability was presented by absorbance value of treated samples compared with absorbance value of same groups 12 hours after loading (%). The survived cells were counted under light microscope and compared with the seeded cells to present the viability (%). (f) G3- and vector-transfected MC3T3-E1 cells were treated with serum free AMEM medium with 2 ng/ml TNF-a, with or without 100 nM SP60025 for 24 hours. All samples were lysed and subjected to immunoblotting with antibodies to pSAPK/JNK, GSK-3β (S9P) and β-actin.

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