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. 2005 Sep;15(9):1189–1197. doi: 10.1101/gr.3873705

Figure 2.

Figure 2.

Determination of the functional PD1 class I core promoter region by mapping of TSSs that contribute to basal promoter activity. (A) HeLa epithelial cells were transiently transfected with wild-type (WT) and class I promoter mutants with out-of-frame ATGs located at positions -6 (ATG-6), -24 (ATG-24), and -42 (ATG-42). The positions of the ATG mutations relative to the determined TSS regions are provided at the bottom of the figure. Constructs with a TAG triplet at positions -6, -24, or -42 served as controls. Data are expressed as relative percentages of acetylation corrected to an internal transfection control, pSV2LUC. Error bars indicate standard error derived from four independent experiments, each performed in triplicate. (B) HeLa cells were transiently transfected with either -416WT or out-of-frame mutant -416ATG-6 reporter constructs. After 48 h, RNA was prepared from half of the sample and used in Northern analysis of CAT reporter RNA (black bars) to quantitate steady-state transcription levels; functional reporter CAT enzyme activity was determined in the other half (gray bars). Results are presented as the relative amount of either CAT reporter activity or RNA, after correcting for transfection efficiency with an internal control pSV2LUC and correcting for RNA loading by tubulin RNA levels, respectively.