SEARCH

SEARCH BY CITATION

As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.

FilenameFormatSizeDescription
emmm201302618.reviewer_comments.pdfPDF document284KReview Process File
emmm201302618-sm-0002-SuppFig-S1.pdfPDF document1575K

Figure S1. Dynamic growth of uterine size and BVs during early pregnancy in mouse. (A) Cross-sectioned uteri from 4.5 to 8.5 dpc stained with hematoxylin and eosin. UL, uterine lumen; E, embryo. (B) Representative images showing CD31+ BVs in the uteri from 4.5 to 8.5 dpc. All scale bars, 500 µm.

Figure S2. Demarcation of BVs from fetus-derived placenta, and distributions of -SMA+ mural cells and SpA in pregnant uteri. (A) Images showing CD31+ BVs and GFP+ embryo inthe uteri at 4.5, 6.5 and 8.5 dpc. Female mice were mated with male GFP+ mice. UL, uterine lumen; E, embryo. (B) Images showing CD31+ BVs and α-SMA+ mural cells at 4.5, 6.5 and 8.5 dpc. All scale bars, 500 µm. SpA, spiral artery.

Figure S3. Role of VEGF-A in decidual angiogenesis and EEVSF, and immunohistochemical localizations of VEGF-A and VEGFR in the pregnant uteri. (A) Images showing CD105+ BVs in the uteri at 8.5 dpc treated with Fc and VEGF-Trap at 6.5 dpc. (B and C) Comparisons of CD105+ BVD (%) in each region, and numbers of different sized VSFs in the CTR. Each group, n = 3. *p < 0.04 versus Fc by one-way ANOVA. (D) Images showing VEGF-A distribution in the uterus at 8.5 dpc. (E) Images showing VEGFR2 distributions in the uterus at 6.5 and 8.5 dpc. Each numbered region (square-dotted line) is magnified and arrayed in right. All scale bars represent 500 µm.

Figure S4. Ovary-secreting E2 does not governs decidual angiogenesis and EEVSF during early postimplantation period. (A) Images comparing CD31+ BVs in the uteri at 6.5 dpc of sham-operated Control and ovariectomized (Ovx) mice supplemented with P4 (Ovx/P4). UL, uterine lumen. Scale bars, 500 µm. (B and C) Comparisons of CD31+ BVD (%) in the ULR, CTR and AMR, and numbers of different sized VSFs in the CTR at 6.5 dpc in Control and Ovx/P4 mice. Each group, n = 4. (D) Images comparing CD31+ BVs in the uteri at 6.5 dpc treated with control and ICI 182780. Scale bars, 500 µm. (E and F) Comparisons of CD31+ BVD (%) in the ULR, CTR and AMR, and numbers of different sized VSFs in the CTR at 6.5 dpc in control and ICI 182780. Each group, n = 4.

Figure S5. Images showing VEGFR3+/Prox-1+/CD31+ lymphatic vessels (arrows) in the myometrium and VEGFR3+/Prox-1/CD31+ VSF in the CTR of endometrium at 8.5 dpc. Scale bars, 100 μm.

Figure S6. Depletion of uNK cells does not reduce expressions of VEGFR2 and VEGFR3 in the vascular sinus folding of pregnant uteri. Images showing VEGFR2 and VEGFR3 expressions in the uteri at 8.5 dpc treated with Fc and anti-NKG2D antibody at 6.5 and 7.5 dpc (twice a day). Each square region (white-dotted line) is magnified in the right side. E, embryo. UL, uterine lumen. Scale bars, 500 μm. Three independent experiments show similar findings.

Figure S7. Normalization of uterine size and BVs, and roles of Ang1, Ang2 and Tie2 during postpartum period. (A and B) Cross-sectioned uteri at PD 0.5, 2.5 and 4.5 stained with hematoxylin and eosin and for CD31+ BVs. Scale bars, 500 µm. Myo, myometrium; EDM, endometrium; UL, uterine lumen. (C) Comparison of CD31+ BVD (%) in the endometrium at PD 2.5 treated with Fc and sTie2. Each group, n = 3. (D) Images showing coverage of α-SMA+ mural cells on CD31+ BVs in the endometrium at PD 2.5 treated with Fc and sTie2. Scale bars, 100 µm. (E) Comparison of coverage of α-SMA+ mural cells on CD31+ BVs (%) in the endometrium at PD 2.5 treated with Fc and sTie2. Each group, n = 3.

Table S1. Effects of various agents on decidual angiogenesis, EEVFS and embryo resorption.

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.