Extended Data Fig. 4: Characterization of the Sel1lVav-knockout mice.
a, b, Expression of Sel1l mRNA (a) or protein (b) in primary control (Ctrl) or Sel1lVav-KO HSCs. n = 5 in a. c, Body weight of Ctrl and Sel1lVav-KO mice at indicated time points, n = 6. d, BM cellularity of Ctrl and Sel1lVav-KO mice at the indicated time points. n = 6. e, Frequency and absolute number of LT-HSCs from 8-week-old Ctrl and Sel1lVav-KO mice. n = 3. f, Colony formation in methylcellulose from equal number of Ctrl or Sel1lVav-KO HSCs. CFU-GM/G/M: colony forming unit granulocytic and monocytic (GM)/ granulocytic (G)/ monocytic (M); CFU-GEMM: colony forming unit granulocytic, erythrocytic, monocytic and megakaryocytic; BFU-E: blast forming unit erythrocytic. The colonies were counted after 12 days of culture. n = 3. g-j, Representative pseudocolor dot plots showing the gating strategy to identify CMP, GMP, MEP, CLP (g, i) and quantification (h, j) in the BM of Ctrl and Sel1lVav-KO mice. n = 6 for each group. k, RBC count in peripheral blood of Ctrl and Sel1lVav-KO mice at indicated age. 8-week-old Ctrl: n = 3; 8-week-old Sel1lVav: n = 7; 50-week-old Crl: n = 7; 50-week-old Sel1lVav: n = 5. l, Kaplan–Meier survival analysis of Ctrl and Sel1lVav-KO mice. n = 20. m, Schematic diagram of the Ki67 assay under BMT condition. n, Cell-cycle analysis of donor and competitor derived HSCs using Ki67 and 7-AAD. n = 3. o, p, Representative FACS plot for apoptosis analysis from freshly isolated control and Sel1lVav-KO HSCs. Sample without Annexin V stain served as negative control. Sample treated by heat shock served as positive control. q, Quantification of apoptotic HSCs in control and Sel1lMx1-KO mice at indicated time points, n = 3. r, Quantification of apoptosis-related genes expression in control and Sel1lVav-KO HSCs. n = 4. s, Quantification of phos-p38, phos-JNK and phos-ERK in control and Sel1lMx1 HSC at indicated time points. n = 3 for all groups except 5 weeks Ctrl (n = 2). t, Percentage of apoptotic HSCs in the BM of 8-week-old Ctrl and Sel1lVav-KO mice. Ctrl: n = 8; Sel1lVav: n = 7. Two-way ANOVA (c, d, h, j, k, q, s), two-tailed Student’s t-tests (a, e, f, n, r, t), or log-rank test (l) were used to calculate P values. Results are shown as mean ± s.d. ns, not significant. Statistical information and unprocessed blots are provided as source data.
