Skip to main content
. 2020 Oct 30;11(10):936. doi: 10.1038/s41419-020-03134-0

Fig. 2. The endogenous expression of miR-140-3p was detected in GECs of BTB and involved in the modulation of BTB permeability in vitro.

Fig. 2

a MiRNA microarray analysis of total RNAs isolated from ECs of BBB and GECs of BTB. Red indicates high relative expression and green indicates low relative expression. b Relative expression of miR-140-3p in ECs and GECs were detected by qRT-PCR. U6 RNA level was used as an endogenous control. Data represent mean ± SD (n = 3, each group). **P < 0.01 vs. ECs group. c MiR-140-3p changed TEER values of GECs. Data represent means ± SD (n = 3, each group). *P < 0.05 vs. agomir-140-3p NC group. #P < 0.05 vs. antagomir-140-3p NC group. &P < 0.05 vs. Control. d MiR-140-3p changed HRP flux of GECs. Data represent means ± SD (n = 3, each group). *P < 0.05 vs. agomir-140-3p NC group. ##P < 0.01 vs. antagomir-140-3p NC group. &P < 0.05 vs. Control. e The relative mRNA expression level of ZAK in GECs was detected by qRT-PCR, and GAPDH was used as an endogenous control. Data represent mean ± SD (n = 3, each group). *P < 0.05 vs. agomir-140-3p NC group. #P < 0.05 and ##P < 0.01 vs. antagomir-140-3p NC group. f The protein expression of ZAK was shown by western blot. g, h The protein expression of NFκB-p65 was shown by western blot. i Western blot analysis of TJ-associated proteins ZO-1, occludin, and claudin-5 in GECs. For panels f–i, GAPDH protein levels were used as an endogenous control. Data represent mean ± SD (n = 3, each group). *P < 0.05 and **P < 0.01 vs. agomir-140-3p NC group. #P < 0.05 and ##P < 0.01 vs. antagomir-140-3p NC group. j Immunofluorescent localization of ZO-1, occludin, and claudin-5 in GECs. ZO-1 (red), occludin (red), and claudin-5 (red) were, respectively, labeled with fluorescent secondary antibody and nuclei were labeled with DAPI. Images were representative of five independent experiments. Scale bar = 20 µm.