Effect of AMC on IPEC-J2 cells proliferation and differentiation in vitro. (A) Cell viability in various concentrations of AMC (0, 0.5, 1.0, 1.5, 2.0, 4.0, 6.0 and 8.0 mg/mL) treatment. (B) Cell viability in various concentrations of LPS (0, 25, 50, 100, 200 and 400 μg/mL) exposure. (C) Cell viability at 2, 4, 6, 8 and 10 h after 100 μg/mL LPS exposure. (D-F) Western blotting measurements of the protein levels of (E) Wnt1 and (F) β-catenin in the IPEC-J2 cells. (G-L) Immunofluorescence images of (H) EdU+ (green), (I) Wnt1 (green), (J) Lgr5+ (red), (K) Muc2 (red), (L) Lys+ (green) and DAPI (blue) staining in IPEC-J2 cells. The mean optical density or cell number was analyzed by Image J software. Statistical analysis was performed using Student’s t-tests to compare differences between the two groups or one-way ANOVA followed by Tukey's post hoc pairwise comparison. Data are presented as the mean ± SD. ns, not significant, *P < 0.05, **P < 0.01, and ***P < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)