To assess cell apoptosis induced by LPS, a TUNEL Cell Apoptosis Assay Kit (C1089, Beyotime, Shanghai, China) was used. Gallbladder mucosal epithelial cells were fixed with paraformaldehyde and then incubated with TUNEL solution for 30 min. After staining with DAPI for 5 min, the cells were imaged using a fluorescence microscope. Image J software was used to analyze cell apoptosis by calculating the number of TUNEL-positive cells.
Apoptosis was further examined using the Annexin V-FITC/PI Cell Apoptosis Detection Kit (C1062L, Beyotime, Shanghai, China). In brief, cells were washed and resuspended with PBS after LPS treatment. Approximately 1 × 105 gallbladder mucosal epithelial cells were stained in the dark with 10 µL of Annexin V-Fluorescein Isothiocyanate (FITC) and 5 µL of Propidium Iodide (PI), followed by apoptosis detection with a flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) (Lu et al. 2022 (link)).
Apoptosis was further examined using the Annexin V-FITC/PI Cell Apoptosis Detection Kit (C1062L, Beyotime, Shanghai, China). In brief, cells were washed and resuspended with PBS after LPS treatment. Approximately 1 × 105 gallbladder mucosal epithelial cells were stained in the dark with 10 µL of Annexin V-Fluorescein Isothiocyanate (FITC) and 5 µL of Propidium Iodide (PI), followed by apoptosis detection with a flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) (Lu et al. 2022 (link)).
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