bec (brain endothelial cells)  (ANGIOGENE PHARMACEUTICALS LIMITED)

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Endothelial cell-specific knockout of Nrf2 impairs BEC homeostasis and reduces endothelial barrier strength (A) qRT-PCR confirms gene expression of exon 5 of Nrf2 is specifically deleted in BECs, but not in astrocytes, microglia or oligodendrocytes in Nfe2l2 ENDO mice. Data are represented as mean ± SE, from left to right p = 0.0007, 0.0995, 0.6903, and 0.7432, n = 4, unpaired t-test. (B) Mouse BECs isolated from Nfe2l2 ENDO mice and Nfe2l2 Flox mice were cultured in vitro and TEER was measured for 24hrs. ∗ p = 0.0005, One-way ANOVA plus Sidak post hoc (n = 8–9). Data are represented as mean ± SE. (C) Scatterplot of RNA-seq analysis showing the influence of Nrf2 specific KO in ECs on BEC transcriptome in basal conditions. Two weeks after tamoxifen-induced recombination, BECs were FACS-sorted from Nfe2l2 ENDO mice and their Nfe2l2 fl/fl littermate’s control mice. Scatterplot was generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 4–6). (D) Volcano plot of mass spectrometry analysis showing the influence of Nrf2 specific KO in ECs on BEC proteome in basal conditions. Two weeks after tamoxifen-induced recombination, BECs were FACS-sorted from Nfe2l2 ENDO mice and their Nfe2l2 fl/fl littermate’s control mice. Highlighted with red and blue dots are the proteins whose expression are significantly increased or decreased respectively ( p < 0.05, n = 4).

Article Snippet: Human Brain Microvascular Endothelial Cells (BECs) were purchase from Innoprot ( P10361 -IM, Elexalde Derio, Spain).

Techniques: Knock-Out, Quantitative RT-PCR, Gene Expression, Isolation, Cell Culture, In Vitro, RNA Sequencing, Control, Generated, Expressing, Mass Spectrometry

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Nrf2 activator RTA-404 modifies the BEC transcriptome under inflammatory conditions Mice were given i.p. injection of either saline or RTA-404 (n = 6–8) daily for 4 days, following which either saline or LPS was given for 24 h. The brain cells were isolated and then sorted with FACS for BECs, RNA extracted and RNA-seq performed. Scatterplots were generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 6–8). (A) Scatterplot of RNA-seq analysis showing the BEC transcriptome modified by peripheral LPS insult compared with saline in Nfe2l2 fl/fl mice (LPS vs. Saline). (B) Scatterplot of RNA-seq analysis showing the LPS-induced BEC transcriptome modified by pre-administration of RTA-404 in Nfe2l2 fl/fl mice (LPS+RTA-404 vs. LPS+saline). (C) IPA analysis identifying activated or inhibited pathways by pre-administration of RTA-404 under conditions of LPS exposure (LPS+RTA-404 vs. LPS+saline). Significant DEGs ( DESeq2 P _adj < 0.05, ∣Log 2 FC∣>1, n = 6–8) with the BEC transcriptome as reference dataset were analyzed to calculate the p-value of overlap and Z score of overall activation/inhibition states of individual pathways. The significant activated (p < 0.05, Z score >2) or inhibited (p < 0.05, Z score<−2) pathways are shown in the bar chart in orange and blue respectively. (D) Volcano plot of mass spectrometry analysis showing the LPS-induced BEC proteome modified by pre-administration of RTA-404 in Nfe2l2 fl/fl mice (LPS+RTA-404 vs. LPS+saline). (E) Scatterplot of RNA-seq analysis showing the LPS-induced BEC transcriptome modified by pre-administration of RTA-404 in Nfe2l2 ENDO mice.

Article Snippet: Human Brain Microvascular Endothelial Cells (BECs) were purchase from Innoprot ( P10361 -IM, Elexalde Derio, Spain).

Techniques: Injection, Saline, Isolation, RNA Sequencing, Generated, Expressing, Modification, Activation Assay, Inhibition, Mass Spectrometry