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. 2019 Sep 12;17(9):e3000421. doi: 10.1371/journal.pbio.3000421

Fig 1. Spatial and temporal contribution of BMDCs to decidua throughout mouse pregnancy.

Fig 1

(A) Biodistribution of BM-derived (GFP+) cells showing preferential recruitment to the pregnant uterus as compared with other body organs. Top panel, PBS nonpregnant control; middle panel, nonpregnant BMT; bottom panel, BMT at E9.5. (B) Engraftment of BMDCs (green) in uteri in nonpregnant state and throughout pregnancy in WT mice receiving BMT from GFP donors. PBS-injected pregnant mouse (E9.5) is shown as negative control, and GFP transgenic mouse is shown as positive control. White arrows indicate the preferential localization of BMDCs to the implantation site. (C) Uterine tissue sections of pregnant mice stained with anti-GFP antibody (brown) showing the localization of BMDCs in decidua during the course of pregnancy. Yellow arrows point to the maternal–fetal interface demarcating the maternal decidua basalis (DB) from fetal placenta (P). Scale bars, 100 μm (top panel) and 50 μm (bottom panel). (D) Representative graphs of flow cytometry of uterine cells demonstrating the temporal changes in hematopoietic (CD45+) and nonhematopoietic (CD45−) GFP+ BMDCs populations during the course of pregnancy. Numbers in each quadrant indicate percentage of cells. (E) Immunofluorescence of uterine tissue sections showing colocalization of PCNA-positive proliferating cells (red) and GFP-positive BMDCs (green) during the course of pregnancy; sections were counterstained with DAPI (blue). Scale bar, 50 μm. (F) Summary of flow cytometry analysis of percentage of GFP+ cells in the uterus during pregnancy (n = 5–7). **p ≤ 0.01 versus E9.5, E13.5, and E17.5. *p ≤ 0.05 versus E9.5. (G) Summary of flow cytometry analysis of percentage of GFP+ and GFP− cells in the uterus that are either CD45+ or CD45− during the course of pregnancy (n = 5–7). *p < 0.01, **p < 0.001. (H) Quantification of proliferating PCNA+GFP+ BMDCs in the uterus during the course of pregnancy (n = 4–7). All bar graphs are mean ± SEM. *p ≤ 0.05. See also S1 Fig. Underlying data are available in S1 Data. BM, bone marrow; BMDC, BM-derived cell; BMT, BM transplant; DB, decidua basalis; E, embryonic day; GFP, green fluorescent protein; NP, nonpregnant; P, fetal placenta; PCNA, proliferating cell nuclear antigen; WT, wild-type.