Fig. 3: NETs promote bEC pyroptosis via the TLR4/NF–κB pathway. | Cell Death Discovery

Fig. 3: NETs promote bEC pyroptosis via the TLR4/NF–κB pathway.

From: NINJ1-mediated plasma membrane rupture of pyroptotic endothelial cells exacerbates blood-brain barrier destruction caused by neutrophil extracellular traps in traumatic brain injury

Fig. 3

A–D Western blotting and quantification of the upstream of TLR4 (NF–κB and p-NF-κB) and pyroptosis-related proteins (GSDMD-N and cleaved caspase-1) in hCMECs (n = 6). NF-κB was used as the loading control for p-NF-κB quantification, and GAPDH was used as the loading control for quantification of pyroptosis-related proteins. E Representative image of double immunofluorescence staining of CD31 (green) and TLR4 (red) in the cortex. Nuclei were stained with DAPI (blue). The white asterisk indicates colocalization of CD31 and TLR4. Scale bar = 50 μm, scale bar on the enlarged images = 10 μm. F–J Western blotting and quantitative analysis of p-NF-κB and pyroptosis-related proteins (GSDMD-N, cleaved caspase-1, and NLRP3) in the cerebrovascular component of the cortex 3 days after TBI. NF-κB was used as the loading control for p-NF-κB quantification, and GAPDH was used as the loading control for the quantification of pyroptosis-related proteins. K, L Representative image of double immunofluorescence staining of CD31 (green) and NLRP3 (red) and quantification of NLRP3 expression in the cortex 3 days after TBI (n = 3). Nuclei were stained with DAPI (blue). Scale bars = 50 μm. Data were presented as the mean ± SD, ns not significant. *p < 0.05, **p < 0.01, ***p < 0.001.

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