MK scRNA-seq. (A) UMAP of scRNA-seq results with identification of different clusters including MKs, T cells, B cells, granulocytes, RBCs, and macrophages. Orange (day-14 BMF), blue (CON). (B) Confirmation of MK cluster by high expression of glycoprotein V (Gp-5), platelet factor 4 (Pf-4), integrin 2 β (Itga2β, Cd41), tubulin β class 1 (Tubb1). (C) GSEA of differentially upregulated gene pathways in day-14 BMF MKs compared with control MKs. (D) Upregulation of genes pertaining to immune activation and antigen presentation and reduction in genes pertaining to platelet function. Individual value plots of Stat1, H2-K1, Nlrc5, Vwf, Igf1r, and Akt3 gene expression of day-14 BMF (orange) and CON (blue). (E) MK subsets. MKs were extracted from whole BM scRNA-seq (panel A), and MK Clusters 0 to 5 were identified. These MKs were classified into 3 groups based on the gene pathways (F). (G) Distribution of BMF and control groups across MK clusters. CA2, calcium²⁺; CON, normal control; iMK, immune megakaryocyte; IL6, interleukin 6; NES, normalized enrichment score.