Fig. 1. Expression of CCR1 protein and mRNA on human EVTs at 9-weeks gestation. (A-E) Serial sections of placental tissue from therapeutic hysterectomy at 9-weeks gestation were doubled-immunostained with anti-CCR1 mAb followed by rhodamine-conjugated secondary antibody plus either FITC-conjugated anti-cytokeratin 7 mAb or FITC-conjugated anti-von Willebrand factor pAb to visualize trophoblasts (A-D) and blood vessels (E), respectively. (C) is a higher magnification of the area indicated in (B). (D,E) The maternal arterial wall is traced with a dashed line. EVTs locating from the cell column (Column) through the trophoblastic shell (Shell) (A,B) and from the trophoblastic shell into the maternal artery (D,E) are shown. CCR1 expression is detected clearly on EVTs in the cell column (B,C) and trophoblastic shell (B), but is scarcely detected on cytotrophoblasts (CT) and syncytiotrophoblast (ST) (C). CCR1 expression is diminished on interstitial trophoblasts (Int TB) (B,C), whereas it is maintained on endovascular trophoblasts (Endov TB) that migrate from the trophoblastic shell into the maternal artery (E). (F) 35-cycle PCR detects a specific band that corresponds to CCR1 in cDNA derived from the microdissected cell columns at 9 weeks of gestation. No band is observed in the negative control in which total RNA from the cell columns was not reverse-transcribed. AV, anchoring villus; IVS, intervillous space; Gl, decidual gland. Scale bars: 200 µm.

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