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Fig. 7 | Journal of Biological Engineering

Fig. 7

From: Long non-coding RNA PVT1 knockdown suppresses fibroblast-like synoviocyte inflammation and induces apoptosis in rheumatoid arthritis through demethylation of sirt6

Fig. 7

PVT1 inhibited sirt6 expression by promoting sirt6 promoter methylation in RA. a, localization of PVT1 in RA-FLSs detected by FISH (× 100); b, the CPG island of sirt6 promoter region analyzed by bioinformatics website MethPrimer; c, the binding site between PVT1 and sirt6 predicted by bioinformatics analysis; d, a bar chart of luciferase activity obtained from the dual-luciferase reporter gene assay; e, strips of sirt6 promoter methylation in RA-FLSs of each group detected by MSP; f, the binding ability of sirt6 promoter to DNA methyltransferase (DNMT1, DNMT3A and DNMT3B) in RA-FLSs measured by CHIP and RT-qPCR; g, the binding ability of PVT1 DNA methyltransferase (DNMT1, DNMT3A and DNMT3B) in RA-FLSs assayed by RIP and RT-qPCR; h, the mRNA expression of sirt6 in RA-FLSs determined by RT-qPCR; i, the protein level of sirt6 in RA-FLSs evaluated by western blot analysis; j, a bar chart of protein level of sirt6 in RA-FLSs; # p < 0.05 vs. the sh-PVT1-NC group; & p < 0.05 vs. the oe-PVT1-NC group; * p < 0.05 vs. the blank group. The results of quantitative analysis were presented in mean ± standard derivation, which compared by one-way analysis of variance; the experimental data was obtained from the average of 3 independent experiments. PVT1, long non-coding RNA plasmacytoma variant translocation 1; Sirt6, sirtuin 6; RA-FLSs, rheumatoid arthritis fibroblast-like synoviocytes; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; RT-qPCR, reverse transcription quantitative polymerase chain reaction; MSP, methylation specific PCR; FISH, fluorescence in situ hybridization; CHIP, chromatin immunoprecipitation; M, methylation; U, unmethylation; MUT, mutant; WT, wild type; DNMT1, DNA methyltransferase 1; DNMT3A, DNA methyltransferase 3A; DNMT3B, DNA methyltransferase 3B

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