Fig. 6

Poor decidual response of hESCs from RIF patients was reversed by increasing Nur77 expression. a hESCs from RIF patients were infected with Ad-Flag-Nur77 or Ad-LacZ (MOI = 20). After 24 h, the cells were treated with 0.5 mM 8-Br-cAMP and 1 μM MPA as indicated for an additional 3 days. dPRL released into the medium was detected by ELFA. ^*** P < 0.001 compared with fertile controls; ^## P < 0.01 compared with RIF hESCs with Ad-LacZ, ANOVA followed by a Student–Newman–Keuls test. (b) and (c) The KLF12 and Nur77 protein levels in hESCs from both fertile women (n = 6) and RIF patients (n = 6) were detected by Western blotting. ^** P < 0.01 compared with fertile controls. d Fluorescein isothiocyanate-labeled phalloidin was used to label actin filaments, and immunofluorescence was used to analyze the morphological transformation of hESCs from RIF patients treated with 0.5 mM 8-Br-cAMP and 1 μM MPA for 3 days after infection with Ad-Flag-Nur77 or Ad-LacZ (MOI = 20). e Attachment of BLSs and their expansion onto hESCs monolayers. hESCs from both fertile women (n = 6) and RIF patients (n = 6) were infected with Ad-Flag-Nur77 or Ad-LacZ (MOI = 20). After 24 h, the cells were treated with 0.5 mM 8-Br-cAMP and 1 μM MPA as indicated for an additional 72 h. Several spheroids were transferred to confluent monolayers of hESCs subjected to the different treatments. Cells were imaged after 24 h, 36 h, and 48 h. f The area of BLS expansion on the RIF hESCs infected with Ad-Flag-Nur77 or Ad-LacZ (MOI = 20) were detected at different time points and is presented relative to that of the Ad-LacZ group (area was set to 1). ^* P < 0.001 compared with Ad-LacZ group at the same time point, repeated measures data ANOVA