Bx53 fl

The immunofluorescence staining was performed as described in previous study [25 (link)]. Briefly, the colon tissue sections (4 μm thick) embedded in paraffin are dewaxed and rehydrated. Then the sections were incubated with primary antibody anti-HOXD10 antibody (1:50, ab138508; Abcam, UK), MUC2 rabbit mAb (1:1,000, A4767; Abclonal, China), Occludin polyclonal antibody (1:1,000, 71-1500; Invitrogen Antibodies, USA), or ZO-1 rabbit pAb (1:1,000, A0659; Abclonal, China) overnight. After washing three times using phosphate buffer saline (PBS; C0221A, Beyotime, China), the sections were treated with biotin-labeled goat anti-rabbit IgG (H+L) with high mole ratio (1:200, A0279; Beyotime, China) for 1 h, followed by incubating with fluorescein diacetate (1:50, 40720ES03; YESEN, China) for another 1 h. Next, the sections were stained with 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (C1002; Beyotime, China) for 30 min. Finally, the images of random six fields were obtained employing a fluorescence microscope (BX53FL, Olympus, Japan).