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. 2015 Sep 14;11(8):1293–1307. doi: 10.1080/15548627.2015.1058474

Figure 7.

Figure 7.

Decrease of promoter methylation and upregulation of MIR4458, MIR4667-5p, and MIR4668-5p in B. pseudomallei-infected A549 cells. (A) Genomic DNA from A549 cells infected with B. pseudomallei for 24 h were subjected to sodium bisulfite sequencing analysis. The frequency of methylated CpGs in the each promoter CpG sites is shown, respectively. Black and white circles correspond to methylated and unmethylated, respectively. (B) The average of methylation at each CpG site within promoters. (C) DNMT1 (DNA MTase) activity assay of nuclear extracts of A549 cells infected with B. pseudomallei for 24 h. (D) Detection of methylation by MS-PCR. A549 cells were treated with DMSO or 5-Aza-CdR (20 μM) for 3 d. M, methylation-specific PCR product; U, unmethylation-specific PCR. (E and F) The expression of MIR4458, MIR4667-5p, and MIR4668-5p was analyzed by qRT-PCR in uninfected or B. pseudomallei-infected A549 cells. A549 cells were treated with 5-Aza-CdR for 3 d at the indicated concentration. Results are represented as mean±SD (*, P < 0.05, **, P < 0.01).