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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Tanshinone IIA alleviates brain damage in a mouse model of neuromyelitis optica spectrum disorder by inducing neutrophil apoptosis

Fig. 6

TSA accelerates human neutrophil apoptosis via caspase-dependent pathways in vitro. Human primary neutrophils and PBMCs (1 × 106 cells/mL/well) were plated in 24-well plates 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 Flow cytometry analysis of cultured neutrophils and PBMCs was performed after APC-Annexin V/PI staining. a Representative flow cytometry plots showing Annexin-V/PI binding (human neutrophils, top; human PBMCs, bottom). b Quantification of the rate of apoptotic cells (human neutrophils, top; human PBMCs, bottom; n = 5, ***p < 0.001). c The cleavage of procaspase-3 was analyzed by western blotting. d Quantification of the relative expression of procaspase 3 and cleaved caspase 3 (n = 5, *p < 0.05, **p < 0.01, ***p < 0.001). The expression of target proteins was normalized to β-actin. The data are representative of three independent experiments. One-way ANOVA followed by Tukey’s multiple comparison tests was performed

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