Fig 1. Transcriptomic and morphological response of MdESF to eutherian decidualizing stimuli.

(A) Experimental strategy and cell type phylogeny of mammalian ESF cell types. ESFs in crown group marsupials and placentals are homologous and descend from a common ancestral therian ESF. In eutherians, DSCs arose from ancestral eutherian ESFs and thus is a sister cell type to eutherian ESFs. (B) Transcriptional response of core regulatory genes involved in eutherian decidualization in MdESF treated for 2 days with either MPA or decidualizing stimuli. Blue dots represent significant differential expression relative to unstimulated MdESF (n = 3, p < 10⁻⁶). Grey dots represent no significant change in expression. Each point represents the mean of three replicates. (C) Gene expression heatmap (mean of three replicates in TPM) of classical decidual markers in HsESF, HsDSC, and MdESF treated with decidualizing stimuli for 2 days. Mean of three replicates is shown. (D) Morphological response of and ROS detection in unstimulated MdESF and MdESF stimulated with either MPA or decidualizing stimuli for 3 days. ROS detection by treatment with H2DCFDA, a general oxidative stress indicator. Scale bars are 10 μm. (E) Differential expression of up-regulated GO categories in MdESF treated with decidualizing stimuli for 2 days relative to unstimulated control (n = 3, adjusted P values for each GO term are shown). Terms related to oxidative stress are indicated in red. Underlying data are provided in S1 Data (Fig 1E) and S2 Data (Fig 1B). Human transcriptome data were previously published in Kin and colleagues [8]. Unedited silhouettes of a monotreme and a marsupial are credited to Sarah Werning and are used under license http://creativecommons.org/licenses/by/3.0/. DSC, decidual stromal cell; ESF, endometrial stromal fibroblast; GO, gene ontology; H2DCFDA, 2′,7′ dichlorodihydrofluorescein diacetate; HsDSC, human decidual stromal cell; HsESF, human endometrial stromal fibroblast; MPA, medroxyprogesterone acetate; ROS, reactive oxygen species; TPM, transcripts per million