The role of Ninjurin-1 deficiency in Jak2V617F clonal hematopoiesis mediated atherosclerosis plaque progression
The role of Ninjurin-1 deficiency in Jak2V617F clonal hematopoiesis mediated atherosclerosis plaque progression
Aortas were mechanically cleaned and then separated into intimal-medial and adventitial layers.  Samples were diced and digested with Liberase, hyaluronidase, and Dnase in media for 45 minutes at 37C. Cells were then washed, blocked with FC block, and hashtaged with TotalSeq-B antibodies and stained for Sytox blue and CD45. Samples were then sorted using flow cytometry (live, CD45+) and subjected to the scRNA-Seq pipeline Ldlr-/- recipients were irradiated, then transplanted with bone marrow from mice with Mx1-Cre driving Jak2V617F +/- Ninj1-/-, 4 weeks later ldlr-/- mice were subjected to a western-type diet (Teklad TD88137) for 12 weeks After 4 weeks of recovery at bone marrow transplantation, mice were subjected to a 12-week Western-type diet (Teklad TD88137) RNA was extracted using the 10x Genomics Chromium Single Cell 3'v4 protocol. Libraries were prepared using the 10x Genomics Chromium Single Cell 3' Reagent Kits v4 according to the manufacturer's instructions. 