DEV015339 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Overexpression of Dkk1. Dkk1 was overexpressed in the peripheral circulation (A) and reproductive tracts (B,C) of pregnant mice. Plasma was obtained by retroorbital phlebotomy from pregnant females receiving injections of empty or Dkk1 ADV (2.5×10⁹ pfu in 0.2 ml saline) on day 1. Oviductal and uterine tissues were collected on day 4. Dkk1 protein migrating as a doublet of 34-38 kDa was detected by western blot (WB) using anti-6xHis antibody (Ab) in the plasma. To examine uterine levels of Dkk1, proteins were first concentrated by immunoprecipitation (IP) using anti-6xhis Ab, and then visualized by WB using anti-Dkk1 Ab. Similar to that in the plasma, Dkk1-His fusion protein was detected as a doublet of 34-38 kDa in day-4 uteri of Dkk1 ADV-treated mice. Immunofluorescence staining in frozen sections further revealed a marked increase in Dkk1 proteins in the oviduct and uterus in females receiving Dkk1 ADV. Liver served as a positive control. Red (Cy3) fluorescence indicates the antigen. Scale bar: 200 µm.

• Supplemental Figure S2 -

  Fig. S2. Cox2 and Pparδ in blastocysts during activation. Cox2 (A) and Pparδ (B) proteins were upregulated in trophectoderm (Tr) cells in activated blastocysts. Representative immunofluorescence staining images depict Cy3-labeled antigens in red, SYTO-13-labeled nuclei in green, and merge in yellow. ICM, inner cell mass; Dor, dormant; Act, activated. Scale bars: 50 µm.