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. 2021 Sep 23;8:91–101. doi: 10.1016/j.aninu.2021.06.010

Fig. 5.

Fig. 5

In-ovo feeding (IOF) lengthens pre-hatch absorptive cell marker expressing regions. (A) RNAscope in-situ hybridization (ISH) of absorptive cell marker peptide transporter 1 (PepT1) at embryonic d 17 (E 17) (pre-IOF) and E 19 in control (non-injected), IOF-NaCl, IOF-Leu and IOF-Gln treated embryos. PepT1 probes were visualized with 3,3′-diaminobenzidine (DAB) and tissues were counterstained with hematoxylin. Dotted lines and arrowheads mark the limit between the PepT1 expressing-region and non PepT1 expressing-region within the villus. (B) Goblet cells were visualized by Alcian Blue acidic mucin staining at E 17 (pre-IOF) and E 19 in control (non-injected), IOF-NaCl, IOF-Leu and IOF-Gln treated embryos. Images were captured at X400 magnifications. Scale bars, 50 μm. (C) Length of non-PepT1 expressing (PepT1-, grey) and PepT1-expressing (PepT1+, brown) segments within each villus at E 17 (pre-IOF) and E 19 in control (non-injected) and IOF-treated embryos. (D) Goblet cell densities at E 17 (pre-IOF) and E 19 in control (non-injected) and IOF-treated embryos. Values are means ± SEM. Different uppercase letters mark significant differences between age/treatment group by Tukey–Kramer HSD, P < 0.05.