Extended Data Fig. 5: Deletion of Piezo1 affects JAM3 and inhibits HSC proliferation.
From: Shear stress governs hematopoietic stem cell fate to promote inflammation-induced aging
a, Schematic view of the LT-HSC lineage tracing. b, Flow cytometry gating strategy and percentages of tdTomato-expressing LT-HSCs in mice treated with Yoda1 (n = 5 mice per group; Yoda1 versus Vehicle P = 0.020). c, Percentages of tdTomato-expressing GMPs in bone marrow (n = 5 mice per group; Yoda1 versus Vehicle P = 0.046). d, Cell counts of tdTomato-expressing neutrophils and Ly6Chi monocytes in blood (n = 5 mice per group; neutrophils: Yoda1 versus Vehicle P = 0.008; Ly6Chi monocytes: Yoda1 versus Vehicle P = 0.018). e, KEGG enrichment analysis of downregulated DEGs in LT-HSCs from Piezo1f/f;Vav mice versus Piezo1f/f controls. The enrichment was evaluated using a two-sided hypergeometric test. f, Heatmap displaying the relative expression levels of DEGs involved in the tight junction pathway. g, Expression levels of genes involved in the tight junction pathway. h, Protein expression of Jam3 in LSK cells (n = 3 mice per group; Piezo1f/f;Vav versus Piezo1f/f P = 0.007). i, Relative Jam3 mRNA expression in LT-HSCs from Piezo1f/f;Vav mice was assessed after passage through the microfluidic device (n = 5 mice per group; Piezo1f/f + Wc 15 μm versus Piezo1f/f + Wc 200 μm P < 0.001, Piezo1f/f;Vav + Wc 15 μm versus Piezo1f/f + Wc 15 μm P < 0.001). j, Percentages of BrdU+ LT-HSCs (n = 5 mice per group; Jam3f/f;Vav-Cre versus WT P = 0.002, Piezo1f/f; Jam3f/f;Vav-Cre versus WT P < 0.001)). The data are representative of two (i) and three (b-d,e-h,j) independent experiments. Data are the mean value ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Statistical significance was determined by two-sided Student’s t test (b-d,h), or two-way ANOVA with Tukey’s post hoc test (i,j).
