Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: TGF- β 1 Induces Interlukin-11 Expression and Pro-Fibrotic Effect by DNA Demethylation in Subconjunctival Fibroblasts

doi: 10.1155/2022/7729827

Figure Lengend Snippet: TGF- β 1 induces DNA demethylation in the IL11 promoter region. After 12 h serum starvation, the HTFs were treated by 10 ng/mL TGF- β 1 for 24 h. A DMEM-alone group was included as a control. The methylation ratios of the 14 CpG sites in the IL11 promoter region were obtained by MassARRAY Platform. (a) DNA fragments from human IL11 promoter region was amplified for the analysis of the methylation level. The fragments were labeled as the promoter region sequence. The location of the promoter region (−1572 bp to −1304 bp) and one CpG island were represented by a blue bar and a green bar, respectively. In light of the reverse complementary strands of these fragments, PCR primer design was performed. The promoter region sequence indicated 269 bp fragments and the CpG sites were counted from 1 to 14 from the 3′- end to the 5′-end. (b) Quantitative methylation levels are shown in a color scale from red to light yellow which represent the methylation level from 0% to 100%, while the grey circle represents missing data. (c) Average methylation levels of the CpG sites in the IL11 promoter region. Data were SD and mean from four independent tests. ∗ P < 0.05, ∗∗ P < 0.01 vs . control group at the same CpG site by one-way ANOVA followed by the Fisher test (C represents Control; T represents TGF- β 1).

Article Snippet: Sequenom MassARRAY platform (CapitalBio, Beijing, China), combined with RNA base-specific cleavage and composed of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, was utilized for the quantitative analysis of IL11 methylation (Gen-Bank Accession Number: NM_000641).

Techniques: Methylation, Amplification, Labeling, Sequencing