Figure 3

Expression of MUC2 and DCLK1 in enteroid 2D monolayers and 3D enteroids stimulated with eqIL-4/IL-13. Relative gene expression of A MUC2 and B DCLK1 transcripts in transwell-grown equine enteroid monolayers basolaterally stimulated with eqIL-4/IL-13 and compared to untreated controls. The gene expression was normalized to the geometric mean for the reference genes HPRT, SDHA and GAPDH and compared to the mean ΔΔCt of the untreated controls. The monolayers were generated from two individual horses and used in two (horse 1) and three (horse 2) separate experiments, giving a sample size of N = 5. C, D Confocal images of equine 3D enteroids cultured in plain growth medium or stimulated with eqIL-4/IL-13 for 48 h before staining with DAPI (blue) and C MUC2 (green) or D DCLK1 (red). Representative images from one horse. E Proportion of cells positive for DCLK1 staining. Mean ± SD of 4 technical replicates. Scale bars = 100 µm for C and 50 µm for D. *P < 0.05, **P < 0.01.