Fig. 3: Motif 1 or a combination of motifs 1 and 2 enhanced expression in a luciferase assay.

HEL cells were transfected with pGL3 plasmids carrying wild-type or mutant motifs 1 and 2 upstream of a luciferase gene driven by the CR1 promoter. Firefly luciferase activity was normalized to a co-transfected Renilla luciferase control and displayed relative to pGL3 carrying only the CR1 promoter. Data represent mean values; error bars indicate standard errors of the mean. The relevant segments of the plasmid constructs are shown on the left of each panel where x indicates the mutant sequence (not to scale). a CR1 motifs 1 and 2 assessed in separate constructs. b CR1 motifs 1 and 2 are assessed as conjoined motifs. CR1-prom, promoter of CR1 (displayed as an open blue box to the left of the diagram); enh, enhancer (displayed as a blue arrow); p1, peak1; m2, peak 2; wt, wild type; mt, mutant. F and R indicate forward and reverse directions respectively relative to the CR1 gene direction. All samples were run in technical triplicates and were repeated in three independent experiments. The ratio between firely and Renilla luciferase luminescence values were calculated, and the means were compared using paired T test (two-sided). Source data are provided as a Source Data file.