Fig. 4

RNA-seq analysis of HSCs from the HU and CON groups. (A-B) Principal component analysis (PCA) of the transcriptional profiles (A) and Pearson’s correlation-based analysis (B) of each sample in the control (CON) and hindlimb unloading (HU) groups. PCA demonstrates the positioning of samples in a three-dimensional coordinate system. (C) Visualization of differentially expressed genes (DEGs) in the HU group compared to the CON group. Red indicates upregulated genes, and blue indicates downregulated genes. (D) Heatmap of the top 10 upregulated and downregulated DEGs. (E) GO enrichment analysis of DEGs, displaying the enriched pathways under three categories: Biological Process (BP), Cellular Component (CC), and Molecular Function (MF). The size of the points represents the number of genes enriched in each pathway, with labels indicating the top five pathways in each category. (F) KEGG enrichment analysis of DEGs. Pathways shared with the GSEA analysis of DEGs are highlighted in red. (G) GSEA enrichment data for the shared KEGG and GSEA pathways. (H-I) GSEA enrichment results visualization for Hematopoietic cell lineage (H) and a manually drawn pathway diagram from KEGG (I). The molecular markers of each cell type are annotated, with color-coded changes in expression between the HU and CON groups: green indicates downregulation, and red indicates upregulation