Skip to main content
. 2014 May 5;9(5):e96005. doi: 10.1371/journal.pone.0096005

Figure 3. The frequency of direct interaction between the kb −148 enhancer and promoter in the KLF4 locus was affected by pitavastatin treatment.

Figure 3

HUVECs were harvested and cultivated as described in the Methods section. (A) The localization of active Pol II obtained by ChIP-seq. The black arrow shows the MEF2C binding site identified by ChIP-seq. (B) A ChIA-PET library was constructed and sequenced. From the TSS of KLF4, 15 PETs originated and 13 of them interacted with a locus −148 kb upstream of the TSS, which result is identical with the MEF2C binding site observed by ChIP-seq and validated by luciferase assay in Figure 2. The numbers in the middle indicate the location on chromosome 9 using the hg19 build program. (C) Quantitative 3C assay. HUVECs were incubated with 1 µM pitavastatin for 4 hours. Primers were designed for analyzing the crosslink frequency of the regions connected with the arches. The relative frequencies were compared between DMSO control (black arch) and statin treatment (red arch). The sequences of the primers are shown in Table S2E in File S1. The data (mean ± SD) is representative of three independent experiments with similar results. Note that the interaction between the TSS and kb −148 was increased by statin treatment.