Fig. 2

Figure 2. CLASP2 is required to maintain HSC activity throughout mouse development (A) Gating strategy to analyze phenotypically defined HSCs (Lin−/lowSCA-1+c-Kit+CD150+CD48−/low) in E14 FLs (n = 3; 8 Clasp2+/+, 7 Clasp2−/−). (B and C) Hematopoietic repopulation analyses after injection of 0.001 ee (B, n = 3) and 0.005 ee (C, n = 3) of E14 Clasp2+/+ and Clasp2−/− FL cells. Red dot, mouse B. (D) Average of CFU-Cs produced weekly per 100 Lin−SCA-1+c-Kit+ (LSK) input cells sorted from Clasp2+/+ and Clasp2−/− E14 FLs and cultured in the LTC-IC culture system for up to 4 weeks. Shown is 1 representative experiment of 2. Error bars, mean ± SD between triplicates. (E and F) Gating strategy to analyze phenotypically defined HSCs in the BM of Clasp2+/+ (top panels) and Clasp2−/− (bottom panels) P8 mice (n = 3) (E) and young adult mice (4.5 weeks old, n = 3) (F). Orange and green dashed lines, highest level of c-Kit and SCA-1 fluorescence, respectively (A, E, and F). Clasp2−/− cells express GFP (as seen in the bottom left plots of A, E, and F; see also STAR Methods). Error bars: mean ± SD (D). ∗∗∗∗p < 0.0001, ∗p < 0.05, Mann-Whitney U test (B and C), Student’s t test (D). FL, fetal liver; BM, bone marrow. See also Figures S1 and S3.