Richardson_Sup Fig 1, 2, and 3

<b>Sup. Fig. 1: RU486 titration in amnion epithelial cells (AECs)</b> <b>A)</b> Documenting morphology of cultured primary AECs under control and RU486 titration conditions. Cells were contoured<b> </b>or puckered after 0.2 µM–0.8 µM RU486 treatment, indicating some form of cell stress. Crystal violet staining was used to show AEC viability (detected by the intensity of crystal violet dye excluded from cells).<b>B)</b> The number of crystal violet positive cells were reduced by half after 48 h with RU486 (≥ 0.2 µM) treatment compared to control (control: 2.8 RIU, 0.01 µM RU486: 2.9 RIU, 0.2 µM RU486: 1.4 RIU, 0.4 µM RU486: 1.4 RIU, 0.6 µM RU486: 1.2 RIU, 0.8 µM RU486: 0.8 RIU). Images were taken at 10X magnification<b>. </b> <b>C-D) </b>The three doses of RU486 that reduced cell viability by one-half after 48 h were added as co-treatments with Dex and cell proliferation and downstream signaling were tested. Co-treatment with Dex+RU486 led to the loss of cells and a stressed phenotype regardless of concentration when compared to control or Dex treatments. Dex+0.4 µM RU486 co-treatment maintained cell viability but was able to antagonize Dex-induced senescence. Due to these two factors, plus the current literature, 0.4 µM RU486 was chosen as the proper dose to antagonize Dex-induced responses. (<b>Proliferation:</b> control: 81.30% RIU, Dex: 78.10% RIU, 0.2 µM RU486: 64.78% RIU, 0.4 µM RU486: 80.36% RIU, 0.6 µM RU486: 57.05% RIU) (<b>Senescence-associated-β-galactosidase:</b> control: 17.47% RIU, Dex: 49.71% RIU, 0.2 µM RU486: 57.47% RIU, 0.4 µM RU486: 25.72% RIU, 0.6 µM RU486: 17.55% RIU). RIU = relative intensity unit.<br> <b> </b> <b>Sup. Fig. 2: Dexamethasone (Dex)- and RU486-induced signaling and cell fate</b> <b>A) </b>11β-hydroxysteroid dehydrogenase-1 (11β-HSD1) was not stimulated by Dex treatment in AECs after 48 h (control 0.28 ± 0.08 RFU, Dex 0.33 ± 0.08 RFU, Dex+RU486 0.37 ± 0.05 RFU) (N = 3). RFU = relative florescence units. <b> B) </b>Treatments with Dex did not induce necrosis in myometrial cells, while Dex+RU468 and RU486 alone increased necrosis when compared with control and Dex treatments (control 8.5±6.9 RFU, Dex 5.7±3.8 RFU, Dex+RU486 24.36±1 RFU, RU486 46.3±16.8 RFU) (Control vs RU486, p=0.005<i>) </i>(Dex vs RU486, p=0.003<i>)</i>. Neither Dex nor RU486 treatment of myometrial cells induced apoptosis (control 36.3±16 RFU, Dex 46.7±19.7 RFU, Dex+RU486 65.1±6 RFU, RU486 40.5±3.6 RFU)<i> </i>(N = 3). RFU = relative florescence units.<br> <br><b>Sup. Fig. 3: Dex does not induce transitions in AECs</b> <br>Morphology of primary AECs under normal cell culture, Dex, and Dex+RU486 conditions<b>.</b> Cells were contoured<b> </b>or puckered after Dex+RU486 but not with Dex alone, indicating some form of cell stress from RU486. Dex nor Dex+RU468 treatment of AECs did not induce cellular transitions measured by E-cadherin/N-cadherin ratio when compared with control (N=3). <br>