As described in pervious study [38 (link)], after treatment, fixed and permeabilized gastric epithelial cell or gastric mucosa tissue slides were incubated with 2% BSA in PBS/0.05% Triton X-100 for 30 min. the slips were incubated with the primary antibody overnight at 4 °C, followed by incubation with Alexa-488- or Alexa 594- conjugated secondary antibodies for 1 h at room temperature. The coverslips were mounted onto glass slides with Prolong Gold Antifade reagent (after staining the nuclei with DAPI), and stained cells were imaged with a Leica TCS SP8 Inverted Fluorescence Microscope (Leica Microsystems). Post-acquisition processing (brightness, opacity, contrast, and color balance) was applied to the entire image and accurately reflected the results of the original image. For statistical analysis, as described in our previous work [39 (link)], 5 areas of each single gastric mucosa section were randomly selected for quantification. The number per square millimeter was counted and analyzed using Leica X image analysis software (Leica, Hamburg, Germany) and ImageJ software (National Institutes of Health, MD, USA).
Free full text: Click here
