Assessment of the effect of SP1 hypomethylation on TLR2 expression. (A, B) 5-AC decreases CpG methylation adjacent to the SP1 binding site within human TLR2 promoter. Genomic DNA from 16HBE14o- cells was either treated with 5-AC for 7 days or untreated (n = 20 for 5-AC-untreated; n = 10 for 5-AC-treated) and then analyzed by sodium bisulfite sequencing. (A) Methylation profiles of the CpG sites (4, 5, 18, 19, 20) adjacent to the SP1 binding sites within TLR2 promoter. (B) The percent of methylated CpGs in each CpG site is shown. (C) Assessment of the effect of Mit A treatment in 5-AC-induced TLR2 up-regulation. 16HBE14o- cells were untreated or treated with 5-AC for 7 days, and then treated with mitA (0.1, 1 μM) for 24 hr or with 100 μM CAPE 3 hr. (D) SP1 expression after 5-AC treatment. 16HBE14o- and CFBE41o- cells were either treated with 5-AC for 7 days or untreated. Total RNA extracted from from both cell lines (in both (C) and (D)) and analyzed by semi-quantitative RT-PCR. TLR2 and SP1 expression was quantified by normalizing to GAPDH and assayed using Image Gauge. Results are representative of three independent experiments.
Furuta et al. BMC Molecular Biology 2008 9:39 doi:10.1186/1471-2199-9-39