Vanillin activates the ASK1-p38 MAPK pathway. CRC cell lines, HT-29/NC, HT-29/shNNMT, SW480/NC and SW480/NNMT, were treated with vanillin and/or SB203580 (SB) for 48 h. (A and B) ASK1-p38 MAPK pathway-related proteins were analyzed by western blot analysis in HT-29/NC and HT-29/shNNMT (A) and SW480/NC and SW480/NNMT (B) cell lines. (C and D) Ratios of phospho-P38 to total P38 (p-P38/P38) were displayed as histograms in HT-29/NC and HT-29/shNNMT (C) and SW480/NC and SW480/NNMT (D) cell lines. (E and F) Ratios of phospho-ASK1 to total ASK1 (p-ASK1/ASK1) are displayed as histograms in HT-29/NC and HT-29/shNNMT (E) and SW480/NC and SW480/NNMT (F) cell lines. The untreated groups of HT-29/NC and SW480/NC were used as the control group, respectively, which was set as 1 in all the histograms. Data are represented as means ± SD, n=3; *P<0.05, **P<0.01, ***P<0.001. Van, vanillin; SB, SB203580; CRC, colorectal cancer; NNMT, nicotinamide N-methyltransferase; ASK1, apoptosis signal regulating kinase 1.