Figure 4

Analyses of mouse Chrna9 promoter in ES cells. (A) 5′-deletion analysis of the mouse Chrna9 promoter region. Progressive deletions of the upstream region are illustrated schematically in the left panel. Each vector was transfected by lipofection into ES cells. At 24 h after transfection, luciferase assays were performed using the cell lysates. Relative luciferase activities are shown. Data are the mean ± SEM values of at least four independent experiments; and *, P < 0.05. (B) Effects of deletion in the T-half site within the Chrna9 promoter region. The constructs used are drawn schematically. Transfection and luciferase assays were performed as described in A. Data are the mean ± SEM values of at least four independent experiments; and *, P < 0.05. (C) Methylation analysis of the promoter region (− 409 to − 190) of Chrna9 gene in undifferentiated ES, differentiated (D5), KUM9 and iPS cells. Each circle denotes cytosine bases in CpG dinucleotides, and filled and open circles represent methylated and unmethylated cytosines, respectively.