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Fig. 7 | Journal of Neuroinflammation

Fig. 7

From: The glucagon-like peptide-1 receptor agonist reduces inflammation and blood-brain barrier breakdown in an astrocyte-dependent manner in experimental stroke

Fig. 7

Blockade of the effects of astrocyte-derived VEGF-A and MMP-9 on the bEnd.3 monolayer protected the TJPs in bEnd.3 cells. a The TEER value of bEnd.3 monolayer cultured with different ACMs for 24 h was assessed. b The ability of NaF to cross the bEnd.3 cell monolayer cultured with different ACMs for 24 h was assessed (n = 6). c Representative western blots for TJPs, including claudin-5, occludin, and ZO-1, in bEnd.3 cells, along with the loading control GAPDH. d The intensity of TJPs was normalized to GAPDH. Quantitative data are presented as the mean ± SD (n = 6). eg Images of immunofluorescence staining for TJPs (claudin5, occludin, and ZO-1) (red) and nuclear staining with DAPI (blue) in bEnd.3 monolayers cultured with different ACMs (n = 6). The arrows indicate the decreased levels of TJPs between bEnd.3 cells. Scale bar = 50 μm. ACM-Medium, cultured with ACM from untreated astrocytes; ACM-OGD+RO, cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 24 h; ACM-OGD+RO+Ex-4, cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 24 h in the presence of Ex-4; ACM-OGD+RO+VI, cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 24 h and the anti-VEGFR2 antibody; ACM-OGD+RO+MI, cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 24 h and batimastat; ACM-Medium+VI, cultured with ACM from untreated astrocytes and the anti-VEGFR2 antibody; ACM-Medium+MI, cultured with ACM from untreated astrocytes and batimastat. All bEnd.3 cell monolayers were pretreated with Ex(9-39) before culture with different ACMs. *P < 0.05 and **P < 0.01 in Fig. 7a, b; *P < 0.05 compared with the ACM-Medium group; #P < 0.05 compared with the ACM-OGD+RO group, $P < 0.05 compared with the ACM-OGD+RO+Ex-4 group, and &P < 0.05 compared with the ACM-OGD+RO+VI group in Fig. 6d, ANOVA plus SNK test (a, b, d)

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