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. 2015 May 12;6(24):20540–20554. doi: 10.18632/oncotarget.4110

Figure 5. Effects of inhibition of the BMP-Smad1/5/8 and Erk1/2 pathways on colon cancer cell proliferation induced by Dragon.

Figure 5

A. and B. Effects of inhibition of the BMP-Smad1/5/8 pathway on Dragon-induced cell proliferation. Stable Dragon over-expression (Dra) and control (pLV) CMT93 cells were incubated for 1 h with and without increasing amounts of the BMP inhibitor LDN193189 (0-50 nM) before the cells were harvested for Western blotting analysis for phospho-Smad1/5/8, Smad1 phospho-Erk1/2, Erk1/2, phospho-p38 and p38 levels, A.. Control (pLV+DMSO), and Dragon overexpressing CMT93 cells in the absence (oeD+DMSO) or presence (oeD+LDN1923189) of LDN193189 at a dose of 10 nM were subjected to CCK-8 proliferation assay at 24, 48 and 72 h after cell seeding B.. Experiments were performed for 3 times independently. C. and D. Effects of inhibition of Erk activity on Dragon-induced cell proliferation. Stable Dragon over-expression (Dra) and control (pLV) CMT93 cells were incubated for 1 h with and without increasing amounts of the Erk inhibitor U0126 (0-10 μM) before the cells were harvested for Western blotting analysis for phospho-Erk and Erk levels C.. Control (pLV+DMSO), and Dragon overexpressing CMT93 cells in the absence (oeD+DMSO) or presence (oeD+U0126) of U0126 at a dose of 0.5 μM were subjected to CCK-8 proliferation assay at 24, 48 and 72 h after cell seeding D.. Experiments were performed for 3 times independently. E. Effects of inhibition of both the Erk1/2 and BMP-Smad1/5/8 pathways on Dragon-induced cell proliferation. Control (pLV+DMSO), and Dragon overexpression CT26.WT or CMT93 cells in the absence (oeD+DMSO) or presence (oeD+U0126+LDN1923189) of U0126 (0.5 μM) and LDN193189 (10 nM) were subjected to CCK-8 proliferation assay at 24, 48 and 72 h after cell seeding. Experiments were performed for 3 times independently. *P < 0.05, oeD+DMSO vs pLV+DMSO.