Figure 3.

Assessment of SP1 binding site interactions with the adjacent CpG#18-20 site. (A) Mutation of the SP1 binding site within TLR2 promoter (-120/+110) decreases basal TLR2 promoter activity in both non-CF and CF epithelial cells. The pGL3-T2P (-120), SP1 binding site-mutant pGL3-T2P (-120) SPm, pGL3-T2P (-12), or pGL3-null plasmids were each transiently transfected into HeLa cells. Luciferase activity is expressed as the percent activity of the pGL3-T2P (-120) construct. (B) Assessment of the effect of mutating the ETS binding site within the TLR2 -120/+110 promoter region. The pGL3-T2P (-120), ETS binding site-mutant pGL3-T2P (-120) ETSm, pGL3-T2P (-12), or pGL3-null plasmids were each transiently transfected into HeLa cells. Luciferase activity is expressed as the percent activity of the pGL3-T2P (-120) construct. (C) Assessment of the effect of mutating the ETS binding site within TLR2 -60/+110 promoter region. The pGL3-T2P (-60), mutant ETS binding site- pGL3-T2P (-60) ETSm construct, the pGL3-T2P (-12) construct, or the pGL3-null plasmid were transiently transfected into HeLa cells. Luciferase activity is expressed as the percent activity of the pGL3-T2P (-60) construct. Data are presented as the mean ± S.E.M. from three independent experiments performed in triplicate. n.s. = not significant as assessed by Student's t test.