Fig. 3

Enhanced KLF12 expression led to a decrease in Nur77 expression. a hESCs were infected with the indicated adenoviruses for 24 h, transfected with NBRE-Luc (300 ng/well), and then treated with 8-Br-cAMP and MPA. After 48 h, luciferase assays were performed, and the data were plotted after normalization to Renilla luciferase activity. ^** P < 0.01 and ^*** P < 0.001 compared with the Ad-LacZ without 8-Br-cAMP and MPA group; ^## P < 0.01 compared with the Ad-LacZ with 8-Br-cAMP and MPA group, ANOVA followed by a Student–Newman–Keuls test. (b) and (c) hESCs were infected with an adenovirus expressing either LacZ or KLF12 for 48 h at the indicated MOI. The KLF12 transcript copy number and protein expression were determined by real-time PCR and Western blotting. ^*** P < 0.001 compared with the Ad-LacZ only-treated group, ANOVA followed by a Student–Newman–Keuls test. (d) and (e) hESCs were infected with Ad-LacZ or Ad-Flag-KLF12 at the indicated MOI for 48 h and then treated with 8-Br-cAMP and MPA for another 2 or 3 days; subsequently, the Nur77 mRNA and protein levels were measured by real-time PCR and Western blotting, respectively. ^** P < 0.01 compared with Ad-LacZ alone; ^# P < 0.05 and ^## P < 0.01 compared with the Ad-LacZ plus 8-Br-cAMP and MPA group, ANOVA followed by a Student–Newman–Keuls test