Figure 5.

High-throughput reporter assays of CREB-bound promoters. A. Experimental design of high-throughput reporter assays. A schematic showing the high-throughput reporter assay experimental design. Promoters are fused to luciferase, transfected in a desired cell line, and stimulated under different conditions. Luciferase activity is measure from uninduced and induced samples and the log2 ratio of these differences is calculated. B. Heatmap of inducible promoter activity. A total of 235 promoters, chosen from CREB ChIP-chip and HaloCHIP-chip data, along with twelve negative control promoters were fused to the luciferase gene, transfected into HeLa cells, and treated with stimulants; forskolin (FSK), PMA, or co-transfected with a transcriptional co-activator, TORC1 +/- FSK. Each box represents the log2 ratio of induced/untreated for each promoter in each condition where the intensity of red is proportional to the strongest induction and the intensity of blue is proportional to the strongest repression. The presence of TORC1 by co-transfection shows the highest number of promoters induced to the highest degree. The box of 12 control promoters at the bottom of the panel are random promoters from the genome and show very little inducible activity in any of the conditions tested.