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. 2020 May 19;11(20):1876–1893. doi: 10.18632/oncotarget.27583

Figure 4. Neuronatin mRNA expression is derepressed with demethylation of the NNAT CpG island by 5-aza-2’-deoxycytidine.

Figure 4

(A) NNAT methylation was quantitated by COBRA analysis in genomic DNA from MNNG/HOS cells and U-2 OS cells following treatment for 72 hours with or without 5aza-dC. Quantitative analysis of the ethidium bromide gel image is shown. (B) Paired OS cell cultures were grown for 72 hours in medium with 5aza-dC (5-Aza) or without (Con). Total RNA was reverse transcribed (RT) and amplified by semi-quantitative endpoint RT-PCR with primers specific for NNAT or SNRPD3 coding sequences. Shown in the ethidium bromide-stained gel image are PCR products from paired samples treated with or without 5aza-dC and with or without RT: Lanes 1,2,3,4 MNNG/HOS; lanes 5,6,7,8 U-2 OS.