Lsr 2 flow cytometry kit

Apoptosis assay was performed with Annexin V-fluorescein isothiocyanate Apoptosis Detection Kit I (BD Pharmingen, San Diego, CA, USA) 48 h following transfection, according to the manufacturer’s instructions. The cell suspension (100 μl) was incubated with 5 μl Annexin V and 5 μl propidium iodide (BD Pharmingen) at room temperature for 10 min. Finally, 400 μl binding buffer was added to each tube and the cells were suspended. The treated cells were analyzed by fluorescence-activated cell sorting using a BD LSR II flow cytometry kit (BD Pharmingen).

EC109 and TE10 cells were treated with 20 μM RGZ for 48 h, and apoptosis rate was measured using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Detection Kit (BD PharMingen, San Diego, CA, USA). Cells were collected, and then washed twice with cold PBS and resuspended in 500 μl of Annexin V-FITC binding buffer. The suspension was incubated with 5 μl of Annexin V-FITC and 5 μl of PI at room temperature in the dark for 10 min. Finally, the apoptosis rate was analyzed by fluorescence-activated cell sorting using a BD LSR II flow cytometry kit (BD PharMingen). The apoptosis rates of cells transfected with specific siRNAs targeting RXRα, PPARg, TLR4, MyD88, ERK, JNK, and p38 or control siRNA were measured using FCM analysis described above. Each experiment was conducted for three times independently.