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Fig. 4 | Epigenetics & Chromatin

Fig. 4

From: Sporadic DUX4 expression in FSHD myocytes is associated with incomplete repression by the PRC2 complex and gain of H3K9 acetylation on the contracted D4Z4 allele

Fig. 4

DUX4 expressing cells have increased H3K9 acetylation and decreased H3K27me3. a Differentiated myocytes from a FSHD1-affected individual were sorted into DUX4 expressing and non-expressing populations using a fluorescent DUX4-target reporter. Antibodies against inhibitory (H3K9me2, H3K27me3) and activating (H3K4me3, H3K9Ac) histone modifications were used to compare differences in modification levels between DUX4 expressing and non-expressing cell populations from the same culture. Percent input normalized to H3 is shown on the Y axis. b The levels of chromatin modifiers, EZH2 (member of the PRC2 complex that methylates H3K27), SUV39H1 (involved in H3K9 methylation) and structural protein CTCF were measured at the permissive contracted D4Z4 array and compared in DUX4 expressing and non-expressing FSHD-affected differentiated myocyte populations. Values shown are the percentage of signal obtained from input chromatin normalized to β-actin. Error bars show standard deviations of 6–12 replicates. Signal was determined by specific PCR amplification of the permissive allele using the LLP primers (see Fig. 1). The presence or absence of RNA polymerase 2 (Pol2) was used as a positive control. Statistical comparison was performed using t test with *p value ≤ 0.05 as being significant

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