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. 2020 Mar 5;10(6):987–1003. doi: 10.1016/j.apsb.2020.02.015

Figure 5.

Figure 5

TIMP1 protects against BBB injury in an MMP-independent manner. (A) Recombinant rAla-TIMP1 protein expressed in eukaryotic cells (293T) was analyzed by SDS-PAGE and Western blots analysis. (B)–(F) HBMECs treated with 1 μg/mL rTIMP1 or rAla-TIMP1 were subjected to hypoxia plus 20 ng/mL IL-1β for 24 h. (B) HBMECs from the indicated treatment groups were subject to transwell permeability assay. Data represent mean ± SEM of six independent experiments (ns, not significant; **P < 0.01). (C) HBMECs from the indicated treatment groups were subject to TEER assay. Data represent mean ± SEM of six independent experiments (ns, not significant; **P < 0.01). (D) Western blot analysis and quantification of indicated proteins from total cell lysates (ns, not significant; *P < 0.05; **P < 0.01). (E) Western blot analysis and quantification of indicated proteins from membrane fragments. Data represent mean ± SEM of three independent experiments (ns, not significant; *P < 0.05; **P < 0.01). (F) Cells were also subjected to IF staining of VE-cadherin, claudin-5, occludin and ZO-1. Scale bars, 10 μm.