Gene expression was measured by real-time polymerase chain reaction (RT-PCR), as previously described (25 (link)). Briefly, total RNA was isolated from colonic tissues using TRIzol (Invitrogen, Carlsbad, CA, USA), and fluoresce were monitored by the SYBR Green detection kit (Thermo Fisher Scientific, Waltham, MA, USA) on a 7900 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The RT-PCR was conducted with primers of the target genes (Supplementary Table 2 ) and the reference gene β-actin, and relative gene expression was calculated by the 2−ΔΔCt method (26 (link)).
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