Fig. 2.

In-ovo feeding (IOF) increases pre-hatch small intestinalepithelial cell quantities. Immunofluorescence of total epithelial cells by 4′,6-diamidino-2-phenylindole (DAPI) staining (blue), SRY-box transcription factor 9 (Sox9) + cells (green), proliferating cell nuclear antigen (PCNA) + cells (red), and merged images at (A) embryonic d 17 (E 17) (pre-IOF) and at (B) E 19 in control (non-injected), IOF-NaCl, IOF-Leu and IOF-Gln treated embryos. Multipotent Sox9+ cells were located from base to mid portion of each villus and co-localized with PCNA+ cells (arrowheads). Progenitor PCNA+/Sox9- cells were located from the mid to upper portion of each villus (arrowhead outlines). Images were captured at X400 magnifications. Scale bars, 50 μm. (C) Quantification of total epithelial cells, multipotent cells and progenitor cells at E 17 (pre-IOF) and E 19 (post-IOF). Values are means + SEM. Different uppercase letters mark significant differences between age/treatment group for each cell type by Tukey–Kramer HSD, P < 0.05.