RNA from snap-frozen tissues was extracted using the NucleoSpin RNA Kit (Macherey-Nagel) according to the manufacturer’s instructions. The intestinal tissue was snap-frozen after removal of the fat tissue, and RNA was extracted using the RNeasy Lipid Tissue Mini Kit (Qiagen). RNA was reverse-transcribed using the High-Capacity cDNA Kit (Applied Biosystems/Thermo Fisher). qPCR analyses were performed on a 7900HT FAST instrument (Applied Biosystems/Thermo Fisher) in triplicate using SYBR Green (Roche). RPLP0 was used as an internal reference gene. Relative expression (RE) was calculated according to the formula REgeneX = 2−(CtgeneX − CtRPLP0) and was expressed as fold change normalized to the control condition. The primer sequences used for qPCR were as follows: RPLP0, 5′-AGATTCGGGATATGCTGTTGG-3′ (forward) and 5′-TCGGGTCCTAGACCAGTGTTC-3′ (reverse); VEGFC, 5′-GGGGGCGAGGTCAAGGCTTTT-3′ (forward) and 5′-CCTGGTATTGAGGGTGGGCTGC-3′ (reverse).

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