Figure 6. Induction of imprinted genes by transient ZAC1 overexpression. mRNA expression of the indicated imprinted genes was measured by qRT-PCR in 22Rv1 cells transfected for 48 h with an expression vector containing cDNA encoding the full-length ZAC1 protein (ZAC1) or the shorter ZAC1 protein isoform (ZAC1Δ). As controls, the empty vector (vector) or the same vector containing the lacZ gene (lacZ) were employed. Transfections were performed in duplicates, PCR for each of which was performed in duplicates. The data presented are mean values from the biological duplicates (bars) of the mRNA expression relative to TBP including standard deviation (error bars). Notably, transfection of the short ZAC1Δ form resulted in higher ZAC1 protein amounts than transfection of the full-length ZAC1 form, as assessed with western blot analysis (insert in the first panel). α-Tubulin was detected as a loading control. The protein samples were loaded in the same sequence as shown in the panel