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. 2020 Jul 23;11(7):560. doi: 10.1038/s41419-020-02791-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Circulating maternal gal-3 evaluated by ELISA in PRINCE cohort of patients (n = 65) during first, second, and third trimester, where some women subsequently development FGR (n = 32). b Western blot analysis of gal-3 expression in placental tissue from normal, healthy human pregnancies (NP) (n = 27) and pregnancies complicated by FGR (n = 19). Densitometry was performed to quantify changes in protein expression using ImageJ. c Representative images of gal-3 and cytokeratin (CK) 7 immunohistochemistry in NP and FGR placentae (n = 14, bar = 100 μm; left panel). The percent staining area of gal-3 and CK7 slides was quantified, and a ratio was calculated by dividing the percent area of gal-3 to CK7 staining in slides from the same placental tissue sample (right panel). d Schematic diagram showing the lack of paternal (mating of Lgals3^−/− (KO) male with Lgals3^+/+ (WT) female, pKO) and maternal (mating of Lgals3^+/+ (WT) male with Lgals3^−/− (KO) female, mKO) gal-3. The offspring is heterogeneous (HT) for gal-3 in both mating combinations (left panel). The abortion rate was calculated on E13 as follows: abortion rate = (fetal resorptions x 100)/total number of implantations (middle panel, n = 6–8). Pictures display whole implantation sites on E13. Arrows point to fetal resorptions (right panel, bar = 1 cm). e The embryo body weight was determined on E13 to identify potential fetal growth restriction (FGR) (n = 6). To assess the fetal development, Theiler stage (TS) analysis was performed. Normally developed embryos on E13 are in TS22 (pKO). FGR embryos are in a lower stage (e.g., TS21 or 21.5) and are characterized by a pinna that is not turned forward and non-separated fingers (arrows; mKO) (bar = 0.25 cm). f Placental efficiency was calculated as fetal/placental weight ratio on E13 (n = 14–21). g Glycogen cells in the spongiotrophoblast of the placenta were stained with periodic acid Schiff (PAS) and counted in 3 squares (1 × 1 mm) per mouse on E13 (n = 14–21). The number of mature uterine natural killer (uNK) cells in the decidua basalis were counted in 3 to 4 squares (1 × 1 mm) per mouse (n = 19–26). In all figures, data are expressed as mean ± SEM. *P < 0.05, ^†P < 0.01, and ^‡P < 0.001 using two-tailed t test.