Annexin 5 fitc flow cytometry assay

Propidium iodide (PI) and Annexin V-FITC-flow cytometry assay (BD Pharmingen, CA) was used to detect the apoptosis in the cells. Cells were treated with 5-fluorouracil (5-FU) for 18–24 h and harvested in PBS. Then the cells were re-suspended in binding buffer (BD Pharmingen, CA), and stained with FITC-conjugated annexin V and PI (Kaijishengwu, China, KGA107). After staining, the cells were analysed by flow cytometry (FACS Calibar; Becton-Dickinson) using Cell Quest software [21 (link)].

A propidium iodide (PI) and annexin V-FITC-flow cytometry assay (BD Pharmingen) was used to detect the apoptosis rate in the cells after SLPI transfection. Briefly, 1 × 10⁶ cells per well were cultured in 6-well plates in the absence of 10% FBS for 48 hours. Adherent cells were detached with 0.25% trypsin without EDTA in 1 × PBS. Cells were harvested in complete RPMI 1640 medium and centrifuged at 1000 rpm for 5 minutes. Each of the cells were washed with 1 × PBS and stained with 50 ug/ml PI and Annexin V-FITC, following the manufacturer’s instructions.

Propidium iodide (PI) and Annexin V-FITC-flow cytometry assay (BD Pharmingen, CA) was used to detect the apoptosis in the cells. Cells were stained with FITC-conjugated annexin V and PI (KeyGEN BioTECH, China, KGA107) according to the manufacturer’s instructions. After staining, the cells were analyzed by flow cytometry (FACS Calibar; Becton-Dickinson) using Cell Quest software.
TUNEL assay was processed using the in situ cell death detection kit (KeyGEN BioTECH, China, KGA7071). Hepatic tissues were stained according to the manufacturer’s instructions. The apoptosis index was captured using a fluorescence microscope (Olympus IX73, Japan).