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

Figure 4. Effects of Dragon inhibition and overexpression on Smad1/5/8, AKT, Erk1/2 and p38 phosphorylation in colon cancer cells.

Figure 4

A. and B. Whole cell lysates from Dragon knockdown (shD) and control CT26.WT and CMT93 cells were used for Western blotting analysis A. for phospho-Smad1/5/8 (p-Smad1/5/8) and total Smad1 (Smad1); phospho-AKT (p-AKT) and total (AKT); phospho-Erk (p-Erk) and total Erk (Erk); and phospho-p38 (p-p38) and total p38 (p38). The Western blots were subjected to densitometric analysis B.. Experiments were performed for 3 times independently. C. and D. Lysates from xenografted tumors from Dragon knockdown (shD) and control CT26.WT and CMT93 cells collected from BALB/c and C57BL/6 mice respectively at day 31 after injections were used for Western blotting analysis for p-Smad1/5/8 and Smad1; p-AKT and AKT; p-Erk and Erk; and p-p38 and p38 C.. The Western blots were subjected to densitometric analysis D.; n = 6). E. Lysates from xenografted tumors from Dragon overexpression (oeDragon) and control CT26.WT cells collected at day 22 after injections in BALB/c mice were used for Western blotting analysis for p-Smad1/5/8 and Smad1; and p-Erk and Erk (right panel). Relative levels of p-Smad1/5/8 to Smad1, and p-Erk to Erk were obtained by densitometric analysis (left panel; n = 6). GAPDH is the loading control. N.S., not significantly different; *P < 0.05; ** P < 0.01.