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Effects of eed loss of function on global levels of histone modifications: (a) after genotyping of the caudal extremity of larvae, 10 to 15 µg of bulk histones from a pool of 10 eed+/+ or eed−/− siblings at 9 dpf were analyzed by Western blot using specific antibodies recognizing H3K27me3, H3K27me2, H3K27me1, H2AK119ub, H3K27ac, H3K9me3 or H4K20me3. After stripping, the membranes were reprobed with an anti-histone H3 as a control; (b) comparison between eed+/+ (red) and eed−/− (green) siblings, of histone modification signals normalized by total histone H3 and expressed relative to the levels found in the wild-type fish. Quantified levels are the mean ± SD of at least three independent histone extractions, except for H3K9me3, where two histone extraction were performed, followed by Western blot analysis. Statistical significance was assessed using a Student t-test; ns, non-significant; *, p < 0.05; **, p < 0.01. (See Supplementary Materials).
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