Fig. 5
KLF12 impaired hESC decidualization in vitro by repressing Nur77 expression. (a) and (b) hESCs were infected with Ad-Flag-KLF12, 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. dPRL released into the medium was detected by ELFA. dPRL mRNA levels were measured by real-time PCR. *** P < 0.01 compared with the 8-Br-cAMP + MPA (−) group; # P < 0.05 and ### P < 0.001 compared with the 8-Br-cAMP + MPA (+) group; and @ P < 0.05 and @@ P < 0.01 compared with the Ad-Flag-KLF12 (MOI = 20) plus 8Br-cAMP + MPA group, ANOVA followed by a Student–Newman–Keuls test. (c) and (d) Attachment of BLSs and their expansion onto hESC monolayers. hESCs were infected with Ad-Flag-KLF12, 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 12 h, 24 h, 36 h, and 48 h. The area of BLS expansion on the hESCs was detected at different time points and is presented relative to that of the Ad-LacZ group (area was set to 1). * P < 0.05 and ** P < 0.01 compared with Ad-LacZ group at the same time point; # P < 0.05 compared with Ad-Flag-KLF12 group at the same time point, repeated measures data ANOVA. e Analysis of blastocyst attachment and expansion. hESCs were infected with Ad-Flag-KLF12, 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 for an additional 72 h. Blastocysts were collected from the uterus of a pregnant mouse in the afternoon at 3.5 days post-coitus (dpc). Several blastocysts were transferred to confluent monolayers of hESCs subjected to the different treatments. Blastocysts were imaged after 24 h and 48 h