Journal: Nature Communications

Article Title: Chronic activation of endothelial MAPK disrupts hematopoiesis via NFKB dependent inflammatory stress reversible by SCGF

doi: 10.1038/s41467-020-14478-8

Figure Lengend Snippet: a , b Immunoblot analysis and quantification demonstrating that expression of IkB-SS in BMECs isolated from CDH5-MAPK mice does not affect ERK1/2 or p65 phosphorylation. Black arrowhead represents endogenous IκBα whereas red arrowhead represents IkB-SS transgene. c , d Representative immunofluorescence images and quantification demonstrating increased levels of nuclear p65 in BMECs derived from CDH5-MAPK mice as compared to controls. Note that expression of IkB-SS in BMECs isolated from CDH5-MAPK mice ( CDH5-MAPK::IkB ) decreases nuclear p65 levels. Each dot within the bar graph represents nuclear p65 staining intensity per individual cell. e , f Heatmap and bar graph demonstrating increased expression of NF-κB-dependent inflammatory genes within BMECs of CDH5-MAPK mice. Crossing CDH5-MAPK with Tie2.IkB-SS mice ( CDH5-MAPK::IkB ) resulted in decreased expression of NF-κB-dependent target genes. Expression of Actb was used for normalization. Dendrograms represent unsupervised hierarchical clustering of the entire dataset ( n = 3 mice/cohort). Color scales represent relative mRNA abundance based on normalized gene expression. Raw data for Heatmaps included in Supplementary Table and Source Data. g Representative immunofluorescence images of femurs intravitally labeled with a vascular-specific CD144/VE-cadherin antibody (red) demonstrating that suppression of NF-κB signaling within endothelial cells of CDH5-MAPK mice resolves their vascular dilatation. Error bars represent sample mean ± SEM. One-way ANOVA for multiple comparisons and Tukey’s correction was performed to determine significance. ** P < 0.01; *** P < 0.001; n.s. P > 0.05.

Article Snippet: IkB-SS expressing lentiviral vector was generated by sub-cloning the sequence of human IκBα super-suppressor (Addgene #15264) into pLVX Puro Vector (Clontech #632164).

Techniques: Western Blot, Expressing, Isolation, Phospho-proteomics, Immunofluorescence, Derivative Assay, Staining, Gene Expression, Labeling

Journal: iScience

Article Title: Transcriptional synergy in human aortic endothelial cells is vulnerable to combination p300/CBP and BET bromodomain inhibition

doi: 10.1016/j.isci.2024.110011

Figure Lengend Snippet:

Article Snippet: 3xFLAG-STAT1 Plasmid , Addgene , 220323.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Isolation, Virus, Sample Prep, SYBR Green Assay, CRISPR, Negative Control, Software, Plasmid Preparation