Claudin 5 Antibody - #AF5216

Fig. 4. Inhibition of protein palmitoylation ameliorates PA induced Sertoli cell dysfunction. (A) Analysis of the palmitoylation levels of proteins extracted from the testes of mice administered or not administered the PA injection, with or without gavage of 2-BP (n = 6 for Control, n = 7 for PA and 2-BP + PA). (B) Analysis of the palmitoylation levels of proteins extracted from primary Sertoli cells, Leydig cells and germ cells, which were treated with or without 0.4 mM PA (n = 3). (C) Inhibition of palmitoylation by 2-BP suppressed PA-induced ER stress in Sertoli cells. Translational expression levels of ER stress-related genes were analyzed using Western blotting (n = 3). (D) ROS detection using DCFH-DA staining. The fluorescence densities were calculated using ImageJ (n = 3). Scale bar: 50 μm. (E) TER detection of primary Sertoli cell barriers. The cells were incubated with PA (PA), with PA combined with 2-BP (2-BP + PA), or with the vehicle (Control) for 3 days after barriers were formed on day 4 (n = 5). (F) FITC-dextran permeability assessment of primary Sertoli cell barriers. The cells were treated PA (PA), with PA combined with 2-BP (2-BP + PA), or with the vehicle (Control) for 24 h after cell barriers were formed (n = 5). (G)Tight junction protein levels were examined by western blotting in TM4 Sertoli cells (n = 3). The relative intensities of bands in western blotting results were quantified by ImageJ and normalized to β-actin levels. Data are presented as mean ± SD. n. s., no significant difference vs. Control group. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control group. #P < 0.05, ##P < 0.01 and ###P < 0.001 vs. PA group.

Figure 1 BBB dysfunction and BEC disruption in post-mortem Alzheimer’s disease brains. (A) Fibrinogen/CD31, Cldn5 and Aβ/Glut1 staining in the cortex and (B) hippocampus of post-mortem health control and patients with Alzheimer’s disease. (C) Quantification of Cldn5 and (D) Glut1 intensity, and (E) Aβ plaques in the cortex and hippocampus of healthy controls and patients with Alzheimer’s disease. Black frames in the low-magnification images indicate the location of the high-magnification images to the right. Arrows indicate Aβ plaques, and arrowheads indicate the fibrinogen leakage and decreased Cldn5 and Glut1 in patients with Alzheimer’s disease. Scale bar = 200 µm in A and B. Data are presented as mean ± SEM, n = 5, *P < 0.05, **P < 0.01.

Fig. 3 Treatment with GTA improved BBB permeability in the peri-infarct area on day 7 after cerebral ischemia. A The representative image of extravagated dextran and IgG in the peri-infarct area. B and C Quantification of dextran and IgG intensity based on A. D The representative image of Evans blue leakage. E Quantification of D. F, Western blotting of tight junction proteins in the peri-infarct area. G–I Quantification of ZO-1, Occludin and Claudin-5 based on F. n = 5 for B and C, n = 4 for E–I. Compared with Sham, *P 

Fig. 4 S@A-NPs treatment ameliorates impairment in BBB of APP/PS1 transgenic mice. (a-f) Coronal hippocampal sections were immunostained for CD31 and ZO1 (a), Claudin 5 (b) or Occludin (c), and DAPI. Percentages of ZO1+ (d), Claudin 5+ (e), and Occludin+ (f) area occupied in the total CD31+ area. (g) Representative images in the cortex and hippocampus coronal sections assessed BBB leakage by 10 KDa and 40 KDa FITC-dextran. (h, i) Percentage of 10 KDa (h) or 40 KDa (i) FITC-dextran+ areas in the cortex and hippocampus coronal sections. Data are presented as mean ± SEM. n = 13–15 slices from 3 mice/group (d, e); n = 14–17 slices from 3 mice/group (f); n = 3 mice/group (h, i); * p 

Fig. 3. Reduction in HG/IL-1β-induced senescence markers and TJ protein degradation in HUVECs by Cur treatment. (A) Typical images of HUVECs stained with SA-β-gal (blue stain indicates senescent cells). The number of SA-β-gal+ cells decreased in the Cur group compared to the HG/IL-1β + DMSO group, whereas it increased after brusatol treatment. Scale bar = 200 μm. (B) Quantification of SA-β-gal+ cells (n = 5). (C) Typical images of Claudin-5 staining (green, TJ protein). The intensity of Claudin-5 increased in the Cur group compared to the HG/IL-1β + DMSO group, which was decreased by brusatol treatment. Scale bar = 100 μm. (D) Quantification of the relative fluorescence intensity of Claudin-5 (n = 5). (E) Analysis of VE-Cadherin, ZO-1, Occludin and Claudin-5 expression in HUVECs by WB (TJ protein). The level of TJ protein was increased in the Cur group compared to the HG/IL-1β + DMSO group, which was decreased by brusatol treatment. (F–I) Quantification of the level of VE-Cadherin, ZO-1, Claudin-5, Occludin (n = 3). (J) Analysis of NRF2, HO-1, P21 and P16 expression in HUVECs by WB. Note: (1) The levels of NRF2 and HO-1 were upregulated in the Cur group compared to the HG/IL-1β + DMSO group, which was reduced by brusatol treatment. (2) The levels of P21 and P16 were decreased in the Cur group compared to the HG/IL-1β + DMSO group, which were increased by brusatol treatment. (K–N) Quantification of NRF2, HO-1, P21, and P16 levels (n = 3). ∗P < 0.05, ∗∗P < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)