Western Blot Analysis of Jejunal Mucosa

We used the jejunal mucosa to prepare the protein samples for western blot assay, as previously described [28 (link)–30 (link)]. The jejunal segments samples were collected from the approximately middle positions in the jejunal tracts, as previously described [28 (link), 31 (link)]. The mid-jejunal segments were rinsed with PBS solution, and then the jejunal mucosa samples were gently scraped off. WCLs of the jejunal mucosa were prepared in RIPA lysis buffer (Sangon Biotech, C500005). The nuclear and cytoplasmic extraction of the intestinal epithelium was conducted using a kit (Thermo Fisher Scientific, 78833). This western blot assay was conducted as previously described [31 (link)]. Primary antibodies included the anti-E-cadherin (Cell Signaling Technology, 14472S), anti-ZO-1 (ABclonal, A11417), anti-Connexin 43 (Proteintech, 26980-1-AP), anti-Lamin B1 (Proteintech, 12987-1-AP), anti-CYP1A1 (Proteintech, 13241-1-AP), anti-AhR (Proteintech, 67785-1-Ig), anti-β-actin (Sigma, A5441), and anti-β-tubulin (Proteintech, 66240-1-Ig). Secondary antibodies included the HRP-conjugated secondary antibodies (Cell Signaling Technology, 7076S and 7074S).