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. 2014 Aug 7;10(8):e1004507. doi: 10.1371/journal.pgen.1004507

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© 2014 Shpargel et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Schematic of MEF TX treatment. (B) Quantitative RT-PCR of indicated genes from Utx^fl/fl;Jmjd3^fl/fl;Cre^ER mouse embryonic fibroblasts (MEFs) left untreated (−TX, light grey) or pre-treated with TX (+TX, black). Significant reductions in Hox expression are indicated (T-test p-value * = 0.05, ** = 0.02, N = 3 samples per treatment). (C) H3K27me3 ChIP of Utx^fl/fl;Jmjd3^fl/fl;Cre^ER MEFs left untreated (green) or pre-treated with TX (red). An IgG control ChIP is illustrated as white bars and H3K27me3 ChIP of ES cells is illustrated as light grey bars. Quantitative PCR of a H3K27me3 negative locus (Slc2a8 promoter, Neg) was utilized for comparison to Hox promoters. Significant H3K27me3 accumulations of MEF +TX relative to −TX are indicated (T-test p-value * = 0.01, ** = 0.04, N = 3 samples per treatment). (D) Quantitative RT-PCR of indicated genes from Utx^+/y;Jmjd3^+/Δ (light grey bars) or Utx^Δ/y;Jmjd3^Δ/Δ MEFs (black bars). Significant reductions in Hox expression are indicated (T-test p-value * = 0.01, ** = 0.003, N = 3 independent MEF lines per treatment). (E) H3K27me3 ChIP of Utx^+/y;Jmjd3^+/Δ (green bars) or Utx^Δ/y;Jmjd3^Δ/Δ MEFs (red bars). An IgG control ChIP is illustrated as white bars and H3K27me3 ChIP of ES cells is illustrated as light grey bars. Quantitative PCR of a H3K27me3 negative locus (Slc2a8 promoter, Neg) was utilized for comparison to promoters of Hox genes and other indicated transcription factors. Significant H3K27me3 accumulations of Utx^Δ/y;Jmjd3^Δ/Δ MEFs relative to Utx^+/y;Jmjd3^+/Δ MEFs are indicated (T-test p-value * = 0.02, N = 3 samples per treatment).