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. Author manuscript; available in PMC: 2019 Sep 6.
Published in final edited form as: Mol Cell. 2018 Aug 2;71(5):848–857.e6. doi: 10.1016/j.molcel.2018.07.005

Figure 2. 6mA is enriched at Polycomb-binding sites in neuronal cells.

Figure 2.

(A) qRT-PCR validated a 70% reduction in DMAD mRNA levels after double-strand siRNA knockdown in BG3C2 cells.(B) Western blot using a DMAD-specific antibody confirmed effective DMAD knockdown in BG3C2 cells.(C) Dot blots using a 6mA-specific antibody confirmed 6mA accumulation in the absence of DMAD in BG3C2 cells.(D) GO analysis showed specific enrichment for neurodevelopment and neuronal functions from downregulated genes carrying intragenic BG3C2 gain-of-6mA regions.(E) Average fold change in 6mA mapped reads versus non-enriched input DNA was calculated for various binned ChIP-chip regions of epigenetic regulators available from the modENCODE database. Average fold change is plotted in Heatmap view. Red (fold change >1) indicates enrichment over input while blue (fold change <1) indicates depletion. 6mA was explicitly enriched at Polycomb protein binding sites. Enrichment and depletion were significant with p-value < 0.001, Welch Two sample t-tests. Data are represented as mean ° SEM.