Fig. 2: CDH5-MAPK mice display systemic and BM-localized inflammation.
a Representative immunofluorescence images of femurs intravitally labeled with a vascular-specific CD144/VE-cadherin antibody (red) demonstrating vascular dilatation in CDH5-MAPK mice. b Quantification of Evan’s Blue Dye (EBD) extravasation (n = 5 mice/cohort). c Representative images of femurs isolated from mice injected with EBD. d Microtiter plate demonstrating intensity of extracted EBD for each sample. Non-injected controls were used to determine baselines. e Heatmap of 242 differentially expressed proteins in plasma of CDH5-MAPK mice identified by proteomic analysis (n = 7 control and n = 8 CDH5-MAPK mice). Color scales represent relative protein abundance reflecting mean fluorescence intensities of SomaLogic aptamer-based ELISA. Raw data included in Supplementary Data 1 and Source Data. f Ingenuity Pathway Analysis of differentially expressed proteins demonstrating that inflammatory responses are over-represented in CDH5-MAPK mice. g, h Immunoblot analysis of BMECs isolated from CDH5-MAPK mice demonstrating that MEK1DD expression in BMECs results in an increase in ERK1/2 and p65 phosphorylation (n = 3 biological replicates per genotype). i, j Representative immunofluorescence images and quantification demonstrating increased levels of nuclear p65 in BMECs derived from CDH5-MAPK mice as compared to controls. Control BMECs treated with TNFα (10 ng/mL for 15 min) were used as a positive control for the assay. Each dot within the bar graph represents nuclear p65 staining intensity per individual cell. Error bars represent sample mean ± SEM. Statistical significance was determined using two-tailed unpaired Student’s t-test. *P ≤ 0.05; ***P < 0.001; n.s.P > 0.05.
