Effects of M2R antagonist methoctramine on cell proliferation. Cells were treated with the indicated concentrations of methoctramine for 72 h. (A and B) Methoctramine inhibited A549 (A) and PC9 (B) cell proliferation in a concentration-dependent manner. Cell proliferation was determined by CCK8 assay. Cells treated with solvent (DMSO) were used as a control. (C and D) Western blot showed that methoctramine decreased MAPK and Akt phosphorylation in A549 (C) and PC9 (D) cells. β-actin was used as loading control for Western blot. (E and F) Quantification of Western blots shown in C and D, respectively. Data were shown as mean±s.e.m. *, P < 0.05; **, P < 0.01; ***, P < 0.001, compared with control.