Fig. 3

ACM from astrocytes treated with Ex-4 ameliorated the OGD+RO-induced destruction of TJPs in bEnd.3 cells. a Representative western blots showing the levels of TJPs, including claudin-5, occludin, and ZO-1 in bEnd.3 cells, along with the loading control GAPDH. b TJP expression was normalized to GAPDH. Quantitative results are presented as the mean ± SD (n = 6). c–e Immunofluorescence staining for the 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 indicated decreased levels of TJPs between bEnd.3 cells. Scale bar = 50 μm. ACM-Medium, cultured with ACM from untreated astrocytes; ACM-Ex4, cultured with ACM from astrocytes treated with Ex-4 for 24 h; ACM-OGD+RO, cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 24 h; ACM-OGD+RO+Ex4, cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 4 h in the presence of 100 nM Ex-4; ACM-OGD+RO+Ex4+Ex(9-39), cultured with ACM from astrocytes treated with OGD for 4 h plus RO for 24 h in the presence of both Ex4 and Ex(9-39); all bEnd.3 cell monolayers were pretreated with Ex(9-39) before culture with different ACMs. *P < 0.05 compared with the ACM-Medium group; ^#P < 0.05 compared with the ACM-Ex4 group; ^$P < 0.05 compared with the ACM-OGD+RO group; and ^&P < 0.05 compared with the ACM-OGD+RO+Ex4group, ANOVA plus SNK test (b)