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. 2022 Dec 6;23(23):15401. doi: 10.3390/ijms232315401

Figure 2.

Figure 2

DNA methylation affects miR-199a expression. (A) Six CSCC lines were treated with 5-AZA for 5 days. Next, RNA was extracted from both treated and control untreated cells and was subjected to qRT-PCR as written in Figure 1. The mean −/+ SD was calculated from at least 6 independent experiments. Statistics were performed using a t-test (* p < 0.05) (** p < 0.01) (B) The effect of treatment with 5-AZA for 5 days on the expression of DNM2 and DNM3OS. (C) In the upper panel drawn a scheme of the genomic region of miR-199a-1 and miR-199a-2 genes and the location of the CpG island that was analyzed. Genomic DNA was extracted from three different PHK lines and three different CSCC cell lines or 3 healthy skin biopsies and 3 CSCC biopsies. The genomic DNA was subjected to bisulfite treatment. The treated DNA was subjected to PCR amplification and cloned into PUC19 (see Materials & Methods). From each sample at least 5 sequencings were performed (gray square represents methylated C residues, the white square represents un-methylated C residue, red represents unclear seq of the residue or missing C residue).