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MOESM6 of Setdb1-mediated H3K9 methylation is enriched on the inactive X and plays a role in its epigenetic silencing

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posted on 18.05.2016, 05:00 by Andrew Keniry, Linden Gearing, Natasha Jansz, Joy Liu, Aliaksei Holik, Peter Hickey, Sarah Kinkel, Darcy Moore, Kelsey Breslin, Kelan Chen, Ruijie Liu, Catherine Phillips, Miha Pakusch, Christine Biben, Julie Sheridan, Benjamin Kile, Catherine Carmichael, Matthew Ritchie, Douglas Hilton, Marnie Blewitt
Additional file 6. Failure of in vivo XCI in Setdb1gt/gt female embryos. a. Genomic location (mm10) of the integration site of the gene trap construct. Arrow heads and long arrow indicate the location of oligonucleotides used to identify the integration b. Domain structure of wild type Setdb1 protein and the predicted gene trap β-geo fusion protein. c. Setdb1 protein expression in wild type and gene trap heterozygote MEFs, as determined by Western blotting (at left), quantified compared to the Actin control (at right). Western blots for H3K9me2, H3K9me3 and H3K27me3 are also shown for the same samples, quantified against tubulin. Bar graphs show quantitation of blots with statistics determined by Student’s one-tailed t-test, *** p < 0.005. (d) qRT-PCR of Setdb1 in primary gene trap heterozygous or wild type MEFs transduced with Nons or Setdb1 hairpins respectively. Relative to Hmbs and normalised to wild type Nons. n = 2, mean ± s.e.m.

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National Health and Medical Research Council

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