Fig. 5

TSA promotes mouse neutrophil apoptosis via caspase-dependent pathways in vitro. Fresh mouse BMNEs (1 × 10⁶ cells/mL/well) were cultured in RPMI 1640 complete medium at 37 °C with 5% CO₂ and stimulated with 100 ng/mL LPS for 21 h in the absence or presence of TSA (1 μM, 5 μM, and 10 μM). a, b Neutrophil apoptosis was assessed by flow cytometry after APC-Annexin V/PI labeling. For each condition, 3 × 10⁴ events were recorded within the neutrophil gate. a Representative flow cytometry plots showing Annexin-V/PI binding. b The apoptosis rate of mouse BMNEs (n = 5, ***p < 0.001). c Western blotting was performed to analyze procaspase 3 (33 kDa) and cleaved caspase 3 (17 kDa and 19 kDa) expression. d Quantification of the relative expression of procaspase 3 and cleaved caspase 3 (n = 5, *p < 0.05, **p < 0.01). β-actin was used as an internal control. The data are representative of three independent experiments. One-way ANOVA followed by Tukey’s multiple comparison tests was performed