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. 2015 Mar 16;16(4):634–643. doi: 10.1080/15384047.2015.1029835

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Figure 3. — Effects of M2R antagonist methoctramine on EMT. Cells were treated with 50 μM pilocarpine and/or methoctramine of indicated concentrations for 72 h. (A and C), Western blot showed that pilocarpine induced EMT, while methoctramine reversed pilocarpine-induced EMT in a concentration-dependent manner in A549 (A) and PC9 (C) cells. (B and D) Western blot showed that methoctramine alone reversed EMT in a concentration-dependent manner in A549 (B) and PC9 (D) cells. Cells treated with solvent (DMSO) were used as a control. β-actin was used as loading control for Western blot. (E) Confocal microscopy images showed membrane expression of E-cadherin (yellow, upper lane) and cytoplasm expression of vimentin (yellow, lower lane). Nuclei were stained with Hoest 33342 (blue). Pilocarpine (10 μM) induced EMT (middle lane), while methoctramine (1 μM) reversed pilocarpine-induced EMT (right lane) in A549 cells.