Annexin 5 fitc pi apoptosis kit

Briefly, cell cycle analysis was assessed with the propidium iodide (YEASEN) by flow cytometry (CytoFLEX, Beckman Coulter, USA) according to standard protocols. For cell apoptosis assay, cells were digested using Trypsin without EDTA. After being washed twice with PBS, cell apoptosis was measured by flow cytometry using Annexin V-FITC/PI Apoptosis Kit (Elabscience, Wuhan, China) according to manufacturer’s instructions. Data were analyzed using the FlowJo 10.6.2 software.

To assess the impact of NaB on MSB-1 apoptosis, a flow cytometry assay was conducted. Cells were treated with or without NaB at a final concentration of 6 mM for 24 h. Subsequently, the cells were suspended and stained with Annexin V-FITC and propidium iodide (PI) according to the instructions provided with the Annexin V-FITC/PI Apoptosis Kit (Elabscience, China), as described by Han et al. (42 (link)). The resulting data were analyzed using Flow Jo V10 software for further interpretation.

Following various transfections, the Kupffer cells were subjected to digestion utilizing 0.25% Trypsin without EDTA. The Annexin V-FITC/PI Apoptosis Kit (Elabscience, E-CK-A211, Wuhan, China) was employed to assess the apoptotic status of the cells, as per the guidelines provided by the manufacturer.

An Annexin V-FITC/PI Apoptosis Kit (Cat. E-CK-A211; Elabscience) was used to detect the apoptosis-inducing effects of doxorubicin in NALM-6 cells. The cells were resuspended in a 500 µL 1× binding buffer with 5 µL Annexin V-FITC and 5 µL PI and incubated at room temperature for 15 min in dark. Apoptosis was detected using flow cytometer (CytoFLEX, Beckman Coulter). The results were analyzed using FlowJo software.

The fraction of cells undergoing cell death in response to shRNA lentivirus was quantified by flow cytometry using an Annexin V-FITC/PI Apoptosis Kit (Elabscience Biotechnology). All procedures were conducted according to the manufacturer’s instructions. Last, the data was recorded by Beckman Coulter CytoFLEX and analyzed using the CytExpert software.

The mouse-immortalized lung epithelium cell line TC-1 was maintained at 37 °C in a 5% CO₂ atmosphere. The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, HyClone) supplemented with 10% foetal bovine serum, penicillin (100 U/mL) and streptomycin (100 mg/mL). For P. multocida infection, cells were infected with log-phase PmCQ2 at an MOI of 1.
For the apoptosis Annexin V-PI staining tests, the harvested cells were stained with Annexin V-FITC and PI according to the manufacturer’s instructions for the Annexin V-FITC/PI Apoptosis Kit (Elabscience, E-CK-A211). Finally, apoptotic cells were quantified by flow cytometry.
For Transwell assays, 8.0 µm pore-size Transwell inserts (Corning, 3422) were used. Briefly, antibiotic-free medium with or without drugs was added to both the upper and bottom chambers, and 1 × 10⁵ cells were seeded in the upper chambers for 16 h. Then, PmCQ2 at an MOI of 1 was added to the upper chambers to incubate with the cells. After 8 h of incubation, the bottom and upper media were collected and diluted to an appropriate concentration to measure the number of translocated bacteria via the plate counting method. Translocation rate = number of translocated bacteria/(number of upper bacteria + number of translocated bacteria) × 100%.