Fig. 2: Zfp90 maintains HSC self-renewal ability and repopulation capacity.

a, b Analysis of apoptotic HSCs (Lin⁻GFP⁺c-Kit⁺Sca-1⁺CD150⁺CD48⁻) in Zfp90^+/+ and Zfp90^−/− mice via Annexin V/PI (a) and active caspase 3 (b) staining 16 weeks after transplantation. c Examination of HSC cell cycle via Ki67 staining in Zfp90^+/+ and Zfp90^−/− mice 16 weeks after transplantation. Proliferative HSCs (Lin⁻GFP⁺c-Kit⁺Sca-1⁺CD150⁺CD48⁻) were defined as Ki67⁺Hoechst⁺ cells. n = 6 for each group. d Zfp90^+/+ and Zfp90^−/− mice were i.p. injected with one dose (200 µg) of BrdU and fed continuously with water containing 800 µg/ml BrdU and 5% glucose for 3 days. Next, BrdU⁺ HSCs (Lin⁻GFP⁺c-Kit⁺Sca-1⁺CD150⁺CD48⁻) were analyzed by FACS. n = 6 for each group. e Zfp90^+/+ and Zfp90^−/− HSCs were cultured in methocult medium for CFC formation assays. CFU-M, CFU-G, CFU-GM, and BFU-E colonies were counted. f 5 × 10⁵ Zfp90^+/+ or Zfp90^−/− BM cells (CD45.2⁺) were transplanted along with 5 × 10⁵ CD45.1⁺ wild-type BM cells into lethally irradiated mouse. The mice were maintained on antibiotic drinking water for 14 days. Chimeras were periodically bled to assess the level of donor-derived blood cells at indicative time points. n = 6 for each group. g Enumeration of donor-derived HSCs per mouse 16 weeks after transplantation over serial transplantation. 1 × 10³ HSC (CD45.2⁺) from Zfp90^+/+ or Zfp90^−/− chimeras were transplanted along with 5 × 10⁵ CD45.1⁺ BM cells into a lethally irradiated CD45¹. recipient. n = 6 for each group. *p<0.05, **p<0.01 and ***p<0.001 by two-tailed Student’s t test. All data presented are shown as the means ± SD collected from three independent experiments