Figure 5

duHMGB1 overexpression in DEF cells favours apoptosis only after LPS activation. A No effect of duHMGB1 overexpression on spontaneous apoptosis. DEF cells were transfected with duHMGB1 plasmid or empty plasmid (control group). Apoptosis was analyzed by flow cytometry using PI (y axis) and FITC-conjugated annexin V (x axis) after additional 48 h of culture. B Effect of duHGMB1 overexpression on apoptosis after LPS stimulation. DEF cells were transfected with duHMGB1 plasmid or empty plasmid (control group), After 24 h, 500 ng/mL LPS was added and the culture was continued for 24 h. The cells were analyzed by flow cytometry for PI (y axis) and FITC-conjugated annexin V (x axis). The total percentages of PI⁻ annexin V⁺ cells (Q3) and PI⁺annexin V⁺ cells (Q2) indicate the apoptosis rate. I, II, IV and V are from a single experiment, which was representative of three separately performed experiments. The bar graphs (III and VI) mean value ± SE of three experiments. Mann–Whitney U test was performed to evaluate the differences. *P < 0.05.