Fig. 5

mcm2 is a direct target of Foxn1. (A) Predicted Foxn1 binding site and ChIP-PCR analysis of the Foxn1 binding to the promoter region of mcm2. The consensus site was marked by colored letters. (B) qRT-PCR analysis of Foxn1 binding to the promoter region of mcm2 in control and foxn1 morphants (mean ± SD, n = 3). (C) The mcm2 promoter and the mcm2p truncated constructs, and the luciferase reporter assay (mean± SD, n = 3). HEK293 cells were cotransfected by the Renilla reporter plasmid and the mcm2 promoter construct together with pCDNA3.3(+)-foxn1. Luciferase assays were determined by using the Dual-Luciferase Reporter Assay System (Promega). 2F/2R stand for a pair of gene specific primers spanning the Foxn1 binding site; CF/CR stand for control primers. The results indicate that the functional consensus Fox binding sites of the mcm2 promoter are positively regulated by Foxn1 in a dose-dependent manner.