Anti connexin 43

Immunohistochemistry was conducted to analyze the jejunal E-Cadherin, ZO-1, Connexin 43, and RegIIIγ expression distribution in mice. These antibodies, including the anti-E-cadherin (Servicebio, GB12083), anti-ZO-1 (Servicebio, GB111402), anti-Connexin 43 (Servicebio, GB12234), and RegIIIγ (Abcam, ab198216) were used in the immunohistochemical analysis. The immunohistochemistry assay was conducted as previously described [24 (link)]. The experiments for negative controls were performed by omitting the primary antibody as previously described [25 (link)–27 (link)]. The mean optical density was analyzed using ImageJ software.

Brain tissues were fixed in paraformaldehyde (4%) and embedded in paraffin and sectioned in 5‐μm slices. The primary antibodies were anti‐glial fibrillary acidic protein (GFAP; Millipore), anti‐heme oxygenase‐1 (HO‐1; Servicebio), and anti‐connexin43 (Servicebio).
Neuronal cell death was measured using co‐staining of TUNEL and NeuN (a specific indicator for neurons). The fixed slices were stained using a Click‐iT TUNEL Imaging Assay kit (Thermo Fisher Scientific) and NeuN (Thermo Fisher Scientific), according to the instructions. The 4′,6‐diamidino‐2‐phenylindole (DAPI) was used as a nuclear specific marker.
The signals in the cortex in the peri‐ischemic region were observed (the focusing area is shown in Figure 1B) and images from three fields in the focusing areas were captured using a microscope (Nikon).