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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Contrasting effect of the latency-reversing agents bryostatin-1 and JQ1 on astrocyte-mediated neuroinflammation and brain neutrophil invasion

Fig. 6

Bryostatin-1 induces NET formation following neutrophil recruitment across a BBB model. ac Neutrophils were either left untreated (a) or treated with bryostatin-1 (25 nM) (b) or PMA (100 nM) (c) for 4 h. In parallel, neutrophils were allowed to transmigrate across a BBB model, with medium containing either IL-8 (1 nM) (d), bryostatin-1 (100 nM) (e), ACM from untreated astrocytes (f), or ACM from bryostatin-1-treated astrocytes (g), placed in the collector. After 4 h, inserts were discarded and DNA was labeled with 1× GreenGlo™ Safe DNA Dye. Cells were then imaged using both FITC (green) and UV (blue) filter sets. h Immunofluorescence microscopy validation of GreenGlo™ Safe DNA Dye labeling by comparison with the commonly used NucBlue® staining. Neutrophils were cultured for 4 h either in medium alone or with bryostatin-1 (25 nM). Next, DNA was labeled either with 1× GreenGlo™ Safe DNA Dye or with NucBlue® and imaged using both FITC and UV filter sets. Stained live cells were then visualized by inverted fluorescence microscope. Original magnification ×200. See also Additional file 2: Figure S2

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