VE-Cadherin Antibody - #AF6265

Fig. 1| Endogenous expression of ZRANB2 and effect of ZRANB2 on biological behaviors of glioma cells. c Protein levels of MMP1, MMP9 and VE-Cadherin regulated by ZRANB2 in U87 and U251 cells. Representative protein expressions and corresponding IDVs of MMP1, MMP9, VE-Cadherin in U87 and U251 are shown; data are presented as mean ± SD (n = 3, each group). *P < 0.05 vs. ZRANB2(−)-NC group, **P < 0.01 vs. ZRANB2(−)-NC group, #P < 0.05 vs. ZRANB2(+)-NC group,##P < 0.01 vs. ZRANB2(+)-NC group.

Fig. 1. Conditioned medium from LPS-stimulated macrophages induces ferroptosis in pulmonary microvascular endothelial cells. (A) Representative immunoblotting and quantification of barrier-associated proteins (ZO-1, VE-Cadherin, and occludin) in the lungs of mice either untreated or subjected to CLP surgery for 6h, 12h, and 24h (n = 3). (B) Representative images and quantification of Evans blue staining in mouse lungs (n = 8). (C) Representative immunofluorescence images of DAPI/CD31/F4/80 staining in mouse lungs. Scale bar = 500 μm. (D) Schematic diagram illustrating the co-culture of septic macrophage-conditioned medium (CM) with human pulmonary microvascular endothelial cells (HPMECs). (E) Representative immunoblotting and quantification of barrier-associated proteins in HPMECs co-cultured with CM, with or without GW4869 treatment (n = 3). (F) Representative images and quantification of PI staining in HPMECs (n = 3). Scale bar = 200 μm. (G, H) Representative images and quantification of liperfluo staining in HPMECs (n = 3). Scale bar = 50 μm. (I, J) C11-BODIPY assay evaluating the lipid ROS levels in HPMECs (n = 3). (K) Representative images for mitochondrial membrane potential of HPMECs (n = 3). Scale bar = 50 μm. (L) Representative immunoblotting and quantification of SLC7A11 and GPX4 in HPMECs (n = 3). Data are presented as mean ± SD.

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.)

Fig. 6. The QL formula preserves BBB integrity in the LPS-induced sepsis model. (A) Schematic of the experimental design, including animal groups, treatment regimens, and timing of EB dye injection. (B) Images depicting EB leakage in brain samples from various groups (n = 5). (C) Quantitative analysis of EB leakage (n = 5). (D) Protein levels of VE-cadherin, MMP-9, occludin, and CAV1 were determined by Western blot, with β-Actin used as a loading control (n = 5). (E–H) Western blot bands for VE-cadherin, MMP-9, occludin, and CAV1 were analyzed semi-quantitatively using ImageJ software. The relative levels of these proteins were calculated based on gray values and normalized to β-Actin (n = 5). (I) CAV1 expression in brain tissues was detected by immunohistochemistry staining (n = 5). Scale bar: 50 μm (upper image), 20 μm (lower image). (J) Collagen IV expression in brain tissues was detected by immunofluorescence staining (n = 3). Scale bar: 100 μm. Data are expressed as mean ± SD.

Fig. 7| MiR-124 inhibits the EGFR signaling pathway and VM formation, that could be rescued by Foxq1 expression. E Immunofluorescence staining of Foxq1, EGFR, VE-cadherin, MMP2 and MMP9 in 5–8F cells that overexpressed miR-124, control or simultaneously Foxq1 and miR-124, respectively; scale bars represent 50μm. qRT-PCR (F) and western blot (G) were used to monitor the expression of EGFR signaling pathway and VM-related genes in 5–8F cells that overexpressed miR-124, control or simultaneously Foxq1 and miR-124, respectively.Data are presented as mean ± SD of three independent experiments.

Fig. 6 |Erlotinib and Nimotuzumab could inhibit Foxq1-induced VM formation and NPC growth and metastasis in vivo.F Statistical results of metastatic nodules of each group; p < 0.001. The expression of related genes in xenograft tumors from each group were monitored by qRT-PCR (G) and western blot (H).