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. 2020 Mar 20;10:5181. doi: 10.1038/s41598-020-62035-6

Figure 1.

Figure 1

Targeted DNA demethylation of the Fgf21 promoter using the dCas9-SunTag and scFv-TET1CD system in Hepa1-6 cells. (a) Schematic figure of the dCas9-SunTag and scFv-TET1CD system. (b) Schematic representation of the promoter region of Fgf21. Open circles and gray boxes indicate CpG sites and PPAR response elements (PPREs), respectively. The region analyzed using bisulfite sequencing (BS) encompassing the transcription start site (TSS) is indicated. Bidirectional arrows indicate primers used for ChIP-qPCR analysis (Fig. 5c). Solid bars indicate selected target gRNAs (gRNA1 and gRNA2). (c) BS analysis of DNA methylation status of the Fgf21 promoter after transient transfection of the system with gRNAs in Hepa1-6 cells. Closed and open circles indicate methylated and unmethylated CpGs, respectively. Representative data of four independent experiments are shown. Statistical analyses were performed using the Mann–Whitney U-test. Data are expressed as mean ± SD. *P < 0.05; N.S., not significant between the denoted pairs.