(A–D) Gross examination and quantification of E12.5 Control and CYP19A1(Tg)Cre; Cdh5fl/fl placentas and weights. Control n = 11, CYP19A1(Tg)Cre; Cdh5fl/fl n = 5. (E–H) Gross examination and …
Excel file containing quantification for embryo weights, placenta weights, trophoblast density, trophoblast migration distance, and fetal labyrinth vasculature in Figure 1.
(A, B) Immunofluorescence staining and quantification of E12.5 Control and CYP19A1(Tg)Cre; Cdh5fl/fl placentas for VE-cadherin (green) and CK8 (red). Yellow arrowheads indicate VE-cadherin+ …
Excel file containing quantification for VE-cadherin expression in trophoblasts, embryo weights, and placenta weights in Figure 1—figure supplement 1.
(A) Sagittal hematoxylin and eosin (H&E) sections from E12.5 Control and CYP19A1(Tg)Cre; Cdh5fl/fl embryos. Boxes show magnified regions of the brain, heart, and liver. Note the thinness of the …
Excel file containing quantification for liver area and myocardial thickness in Figure 1—figure supplement 2.
(A) Immunofluorescence staining of E12.5 Control and CYP19A1(Tg)Cre; Cdh5fl/fl brains for VE-cadherin (green) and Endomucin (red) at sites of hemorrhage. Images in gray scale are VE-cadherin alone. …
Excel file containing quantification for VE-cadherin expression in various organs in Figure 1—figure supplement 3.
(A) Immunofluorescence staining of E12.5 Control and CYP19A1(Tg)Cre; Cdh5fl/fl placentas for Endomucin (green) and alpha-smooth muscle actin (αSMA) (red). White arrowheads indicate αSMA+ cells. …
Excel file containing quantification for smooth muscle cells per spiral artery, spiral artery diameter, and trophoblast-endothelial cell displacement in Figure 2.
(A) Schematic of workflow for Doppler ultrasound of pregnant dams. The embryo, UA/UV (yellow arrow), and placenta (dotted white outline) are labeled. (B) Representative umbilical artery Doppler …
Excel file containing quantification for ultrasound studies (resistance index, pulsatility index, fetal heart rate) in Figure 3.
(A) Schematic of bulk RNA-sequencing (RNA-seq) workflow on deciduas from Control (WT) and CYP19A1(Tg)Cre; Cdh5fl/fl (KO) E12.5 placentas. (B) Heatmap of the top 100 differentially regulated genes …
(A) Mmp15 mRNA gene expression in control and CYP19A1(Tg)Cre; Cdh5fl/fl placentas from RNA-sequencing (RNA-seq). (B) Immunofluorescence staining for Endomucin (green), CK8 (red), and MMP15 (gray) in …
Excel file containing quantification for Mmp15 gene expression, laminin expression in the decidua, and vinculin expression in trophoblasts in Figure 4—figure supplement 1.
(A) Immunofluorescence staining for DBA (a lectin that specifically binds uterine natural killer (uNK) cells, gray), vimentin (green), and cleaved caspase-3 (red).
Dotted line demarcates the border of the junctional zone and decidua. Scale bars = 50 μm. (B, C) Quantification of DBA+ cell density and cleaved caspase-3+ cell density in the decidua. (D) Gene ontology (GO) analysis of upregulated biological process terms related to NK cells. Statistical analysis was performed using two-tailed, unpaired Welch’s t-test. Data are shown as means ± SD.
Excel file containing quantification for uterine natural killer cell density and apoptotic cell density in Figure 4—figure supplement 2.
Cdh5fl/fl male mice were crossed with CYP19A1(Tg)Cre; Cdh5fl/+ females to generate litters with mixed genotypes. The expected percentage is listed under the genotype label. The observed number of …
CYP19A1(Tg)Cre; Cdh5fl/fl (25%) | CYP19A1(Tg)Cre; Cdh5fl/+ (25%) | Cdh5fl/fl (25%) | Cdh5fl/+ (25%) | Total (100%) | Fisher Exact Test | |
---|---|---|---|---|---|---|
E10.5-E12.5 | 13* (26%) | 13 (26%) | 12 (24%) | 12 (24%) | 50 (100%) | P=1.0000 |
E14.5-E16.5 | 1 (2.9%) | 13 (38.2%) | 12 (35.3%) | 8 (23.5%) | 34 (100%) | P=0.0033 |
P21 | 1 (2.2%) | 11 (24.4%) | 15 (33.3%) | 18 (40.0%) | 45 (100%) | P=0.0333 |
One embryo at E12.5 was dead.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (Mus musculcus) | CYP19A1(Tg)-Cre | Wenzel and Leone, 2007 | ||
Genetic reagent (Mus musculcus) | Cdh5 flox | Yang et al., 2019 | ||
Antibody | Anti-Endomucin (goat polyclonal) | R&D | AF4666 | IF(1:400) |
Antibody | Anti-Endomucin (rat monoclonal) | Abcam | ab106100 | IF(1:300) |
Antibody | Anti-TER119 (rat monoclonal) | Abcam | ab91113 | IF(1:300) |
Antibody | Anti-VE-cadherin (goat polycloncal) | R&D | AF1002 | IF(1:200) |
Antibody | Anti-CK8 (rabbit monoclonal) | Abcam | ab53280 | IF(1:300) |
Antibody | Anti-CK8 (rat monoclonal) | DSHB | TROMA-1 | IF(1:400) |
Antibody | Anti-αSMA-Cy3 (mouse monoclonal) | Sigma | C6198 | IF(1:300) |
Antibody | Anti-Cleaved Caspase-3 (rabbit polyclonal) | Millipore Sigma | AB3623 | IF(1:100) |
Antibody | Anti-MMP15 (rabbit polyclonal) | Thermo Fisher Scientific | PA5-13184 | IF(1:200) |
Antibody | Anti-Laminin (rabbit polyclonal) | Sigma | L9393 | IF(1:200) |
Antibody | Anti-Vimentin (goat polyclonal) | R&D | AF2105 | IF(1:300) |
Antibody | Anti-Vinculin (mouse monoclonal) | Sigma | V9131 | IF(1:200) |
Commercial assay or kit | Direct-zol RNA Miniprep Kits | Zymo Research | R2053 | |
Software, algorithm | ImageJ | NIH, Bethesda, MD, USA | RRID:SCR_003070 | |
Software, algorithm | GraphPad Prism | GraphPad | RRID:SCR_002798 | |
Software, algorithm | Picard v2.17.11 | Picard | RRID:SCR_006525 | |
Other | DBA-Biotin | Vector Labs | B-1035 | (1:500) |