Journal: bioRxiv

Article Title: PPARG directs trophoblast cell fate and establishment of the uterine-placental interface

doi: 10.1101/2025.10.08.681177

Figure Lengend Snippet: A) In situ hybridization showing Prl7b1 transcripts (cyan) in gestation day ( gd ) 18.5 Pparg f/f and Pparg d/d placentation sites. B ) Immunohistochemical localization of KRT8 (cyan) in gd 18.5 Pparg f/f and Pparg d/d placentation sites. C ) RT-qPCR analysis of Prl7b1, Prl5a1, and Krt8 expression in the uterine-placental interface of gd 18.5 Pparg f/f and Pparg d/d placentation sites (n=6). D ) Quantification of trophoblast cell invasion area (magenta dotted line) (n=4). E ) Immunohistochemical localization of perforin ( PRF1 ), a natural killer ( NK ) cell-associated protein, (magenta) and KRT8 (cyan). F ) RT-qPCR analysis of NK cell-associated transcripts ( Prf1 and Ncr1 ) in the uterine-placental interface of gd 18.5 Pparg f/f and Pparg d/d placentation sites (n=6). G ) Immunohistochemical localization of smooth muscle actin ( SMA , magenta) and KRT8 (cyan) in the uterine-placental interface of gd 18.5 Pparg f/f and Pparg d/d placentation sites. H ) RT-qPCR analysis of Sma transcript in the uterine-placental interface of gd 18.5 Pparg f/f and Pparg d/d placentation sites (n=6). Scale bars for images in panels A, B, E and G=500 μm. Data are presented as mean ± standard error of the mean. Each data point represents a distinct uterine-placental interface sample obtained from four to six pregnancies. Statistical analyses were performed using unpaired t -tests: * p <0.05, **p<0.01, *** p <0.001, **** p <0.0001. Abbreviations: UPI, uterine-placental interface; JZ, junctional zone; LZ, labyrinth zone; SpA, spiral artery.

Article Snippet: Tissue sections were blocked with 10% goat serum (50062Z, Thermo Fisher) and incubated overnight at 4°C with primary antibodies against vimentin (1:300, sc-6260, Santa Cruz Biotechnology), pan-cytokeratin (1:100, F3418, Sigma-Aldrich), perforin (1:300, TP251, Amsbio), or PPARG (human: 1:100, sc-7273, Santa Cruz Biotechnology; rat: 1:100, 2435S, Cell Signaling Technology).

Techniques: In Situ Hybridization, Immunohistochemical staining, Quantitative RT-PCR, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Interleukin-15 deficient rats have reduced osteopontin at the maternal-fetal interface

doi: 10.3389/fcell.2023.1079164

Figure Lengend Snippet: Reduced OPN in the decidua of IL15Δ/Δ rats. (A) Relative expression of Spp1 mRNA in WT versus IL15Δ/Δ (mutant) implantation sites on GD 9.5 ( N ≥ 13 implantation sites from at least 5 dams per group). (B) Images of OPN immunohistochemistry in WT versus IL15Δ/Δ implantation sites. Scale bar, 250 µm. The boxes show the approximate location of high magnification images in (C) for the decidua basalis (DB), near the ectoplacental cone (EPC), and at the periphery with uterine glands. (C) Higher magnification images showing OPN localization in the DB, near the EPC, and uterine glands of WT and IL15Δ/Δ implantation sites on GD 9.5. The bottom panels show images captured following immunohistochemistry of WT implantation sites using a non-specific mouse IgG 1 in place of OPN. Scale bar, 50 µm. (D) Quantification of OPN-positive cells in DB, EPC, and uterine glands from WT and IL15Δ/Δ implantation sites ( N ≥ 5 implantation sites from different dams per group). (E) Immunohistochemistry for PRF (green) and OPN (red) in a representative WT GD 9.5 implantation site. Images were taken of the DB and near the EPC. Please note that PRF and OPN frequently colocalize. Scale bars represent 50 μm. Panel (A) was analyzed using a linear mixed model followed by Tukey’s post hoc test; other experiments were analyzed using Student’s t -tests. Results represent means ± SEM. Data significantly different from controls ( p < 0.05) are indicated by an asterisk (*).

Article Snippet: Cell smears were permeabilized using 0.3% Triton-X and 1% BSA in PBS, blocked with 10% normal goat serum (ThermoFisher Scientific), and immersed in primary antibodies specific for OPN (0.5 μg/mL, sc-21742, Santa Cruz Biotechnology) and PRF (2.5 μg/mL, TP251, Torrey Pines Biolabs) overnight.

Techniques: Expressing, Mutagenesis, Immunohistochemistry

Journal: Frontiers in Cell and Developmental Biology

Article Title: Interleukin-15 deficient rats have reduced osteopontin at the maternal-fetal interface

doi: 10.3389/fcell.2023.1079164

Figure Lengend Snippet: Isolation and enrichment of Spp1 -expressing NK cells from the rat decidua. (A) Schematic depiction of experimental protocol. (B) Relative gene expression of Ccl5 , Klrb1f , Prf1 , Spp1 , and Vim in adherent and suspended cells isolated from GD 9.5 decidua and cocultured for 24 h. Experiments were performed 6 times using different dams. (C) Non-adherent cells isolated from GD 9.5 decidua express both PRF and OPN. Arrowheads indicate a representative cell positive for both PRF and OPN staining. Scale bars, 50 μm. Results represent means ± SEM. Data significantly different from controls ( p < 0.05; Student’s t -test) are indicated by an asterisk (*).

Article Snippet: Cell smears were permeabilized using 0.3% Triton-X and 1% BSA in PBS, blocked with 10% normal goat serum (ThermoFisher Scientific), and immersed in primary antibodies specific for OPN (0.5 μg/mL, sc-21742, Santa Cruz Biotechnology) and PRF (2.5 μg/mL, TP251, Torrey Pines Biolabs) overnight.

Techniques: Isolation, Expressing, Staining