Journal: Nature Communications

Article Title: Post-myocardial infarction heart failure dysregulates the bone vascular niche

doi: 10.1038/s41467-021-24045-4

Figure Lengend Snippet: a scRNA-seq of mice bone marrow vascular niche 7 (d7) and 28 days (d28) after myocardial infarction as well as of controls (d0). Clustered cells from the three-time points are displayed in t-SNE plots. Emcn expressing cells are colored. b -left Representation of the percentage of Emcn expressing cells of the lineage depleted, CD31 expressing population. b -right Emcn and ( c ) Il1b and Myc average expression. d Representation of the ten most significant upregulated terms at d7 revealed by gene ontology analysis when comparing d0 to d7. e Immunostaining of longitudinal femur sections at d1. IL-1β immuno-signal is enriched in type H vessels one day after the ischemic insult. f -up RT-qPCR analysis of MYC expression in HUVEC cells after 1 h IL-1β treatment. N = 4. Data are shown as mean ± SEM. P -value was calculated by a two-tailed Mann–Whitney test. f -down Analysis of MYC activation in HUVEC cells after 1 h IL-1β treatment. N = 3 independent experiments with two technical replicates. Data are shown as fold-change relative to control. g Analysis of Caspase1 activation in HUVECs after 1 h IL-1β treatment or nigericin as a positive control. Left, Gating strategy. Right, quantification. N = 4. Data are shown as mean ± SEM. P -value was calculated by a two-tailed Mann–Whitney test. h Analysis of human MYC expression in isolated liver endothelial cells by RT-qPCR. N = 2 for both groups. Data are shown as mean ± SEM. i Immunostaining of longitudinal sections through the femur. The length of type H vessels (indicated by the dashed line, endomucin in red, CD31 in green, and DAPI in blue) is reduced in Myc EC-OE mice when compared to controls. Tamoxifen was given at 8 weeks and analysis performed 4 weeks later, N = 6 for both groups. Data are shown as mean ± SEM. P -value was calculated by unpaired, two-tailed Student’s t -test.

Article Snippet: HUVEC cells were seeded in 6 well cell culture plates (657160; Greiner Bio-One) and stimulated after 24 h with Nigericin (10 μM; N7143; Sigma Aldrich) or IL-1β (100 ng/mL; 201-LB; R&D Systems) for 3 h at 37 °C.

Techniques: Expressing, Immunostaining, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Activation Assay, Control, Positive Control, Isolation