Fig. 2

Gene profiles of trophoblast outgrowths after 6 days of culture from in vivo-derived dams. All data are presented as mean ± SEM, analysed by Student’s t test. White (male), black (female). a Representative image of day 6 cultured trophoblast outgrowth from E5 stained with trophoblast-specific marker pan-cytokeratin (red) and DAPI (blue). b Trophoblast outgrowth area over time from time 0 (E5-derived embryo). Assessed at day 6 of culture were c number of trophoblasts (nuclear area > 200um²), d DNA content relative to 2N ICM cells, e parietal trophoblast giant cell (P-TGC) number (nuclear area > 1000 um²), f trophoblast nuclear area, g P-TGC nuclear area and h–k expression profiles. h-k gene expression profiles displayed show the gene of interest relative to the geometric mean of 3 housekeeper genes (Rplp0, Rpl13a, 18S). Expression of Prl3d1 k showed unequal variance and was analysed by Mann-Whitney non-parametric test. Data in b show litter averages from N = 4 litters, 2–9/sex/litter. Cell counts and DNA content (c–g) show litter averages of N = 4 litters, 1–3/sex/litter. QPCR (h–k) show N = 8/sex from 4 litters