FIG. 4.

The −298 ATTA site is sufficient to confer transcriptional activation by LHX3. Transient transfections were performed in αT3-1 cells using a luciferase reporter containing four tandem copies of either the WT (−298 multi) or the mutant −298 ATTA (−298 mut multi) element cloned upstream of a minimal −81 thymidine-kinase promoter. Either an empty overexpression plasmid or one containing the LHX3 cDNA were cotransfected with the multimerized reporter, and reporter gene expression was assayed. Data are the means ± sem of at least three independent experiments, each performed in triplicate, and were normalized as described in Materials and Methods. A one-way ANOVA followed by Student’s t test was performed to analyze statistical significance. *, Significant difference from cells cotransfected with empty vector (P ≤ 0.05).