Fig 2: Angiogenesis on cyclophosphamide-(CTX)-treated ovaries after adipose-derived mesenchymal stem cell (A-MSC) and tail fibroblast transplantation. (a) Immunohistochemistry (IHC) of CD34 on rat ovaries. Left, is the cell or saline administration; right, non-injected control. Fibroblast: tail fibroblast cells; adipose: A-MSCs. Scale bar=200 µm. (b) The number of CD34-positive cells in corpus lutea (CL) in ovaries. The number of CD34-positive cells per 1 mm2 of CL was examined. Significant differences between each character is represented by a, b and c (P<0.05). Fibroblast: tail fibroblast cells; adipose: A-MSCs.

Fig 3: Nicotine induces HSC quiescence after transplantation.a Scheme illustrating experimental design. Irradiated WT CD45.2 recipients were transplanted with 2 × 106 BM nucleated cells from CD45.1 mice and treated with 65 µg/ml nicotine or vehicle in their drinking water over 7 weeks. b, c Circulating donor-derived (CD45.1+) haematopoietic cells (b, P = 0.03) and white blood cells (c, P = 0.006,0.04) at specified time after transplantation (N = 3). d Donor-derived B cells (P = 0.007), T cells (P < 0.0001) and myeloid cells (P = 0.009) from mice treated with nicotine or vehicle at 7 weeks (N = 3). e Cell cycle profiles of donor-derived lin-sca1+ckit+ (LSK) CD34-flt3- HSCs isolated from the endosteal or central BM 7 weeks after transplantation and chronic treatment with nicotine or vehicle (N = 3;P = 0.005,0.04). (b–e) Data are mean of biological replicates ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. (b–e) ANOVA and Sidak’s comparisons.

Fig 4: Nicotinic receptor blockade reduces HSC quiescence after transplantation.a Scheme illustrating the treatment of sub-lethally irradiated WT CD45.2 recipients of 2 × 106 BM nucleated cells from CD45.1 mice, treated with Hexamethonium bromide (20 mg/kg/d over 30d, i.p.) or vehicle. b Cell cycle profiles of donor-derived (CD45.1+) lin-sca1+ckit+ (LSK) CD34-flt3- HSCs isolated from the endosteal or central BM 30 days after transplantation and chronic treatment with hexamethonium bromide or vehicle (N = 5;P = 0.002,005). Data are mean of biological replicates ± SEM. **P < 0.01. ANOVA and Sidak’s comparisons. c Representative flow cytometry plots showing HSCs in different phases of the cell cycle. The frequencies of the gated populations are indicated.