ANGPTL2 supports HSC function by sustaining the level of G0S2. (A) Relative mRNA expression of potential candidates (Rora, Rab20, Pik3r2, Isg15, and G0s2) in LT-HSCs purified from Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl mice as measured by quantitative RT-PCR (n = 3). (B) Relative mRNA expression of G0s2 in total BM cells, Lin⁻ cells, MPPs, short-term HSCs, and LT-HSCs from Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl mice as measured by qRT-PCR (n = 3). (C) Nucleolin expression (red) in LT-HSCs from Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl mice was examined by immunofluorescence staining. Representative images are shown on the left. A total of 17 to 19 LT-HSCs were evaluated in the Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl groups (right, n = 3). (D) Representative flow cytometric analyses for the repopulation of HSCs from Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl mice with/without G0s2 overexpression (GFP⁺ cells) 16 weeks after transplantation. (E) The quantification data of the donor contribution at the indicated time points after transplantation are shown (n = 5). (F) The cell-cycle status of LT-HSCs from Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl mice with/without G0s2 overexpression was determined by staining with Hoechst 33342 and pyronin Y. The percentages of the G₀ fraction are shown (n = 3). (G) Nucleolin localization in LT-HSCs from Angptl2^fl/fl and Cdh5-Cre; Angptl2^fl/fl mice with/without G0s2 overexpression was examined by immunofluorescence staining. A total of 45 to 55 LT-HSCs were analyzed. *P < .05; **P < .01; ***P < .001.