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Fig. 3 | Epigenetics & Chromatin

Fig. 3

From: JMJD1B, a novel player in histone H3 and H4 processing to ensure genome stability

Fig. 3

siJMJD1B cells accumulate histones H3 and H4 at early steps of the maturation cascade associated with tNASP/Hsp90/Hsc70. a Western blot of fractions derived from the sizing exclusion chromatography S200, loaded with 0.2 mg of cytosolic extracts derived from siControl or siJMJD1B HeLa cells. The corresponding molecular size of each fraction is specified at the top of the western blot. b Western blot of samples derived from immunoprecipitation assays using 1.5 mg of cytosolic extracts derived from siControl or siJMJD1B HeLa cells that express exogenously low levels of Flag-H3.1 protein. Immunoprecipitation was performed using antibodies against Flag. Input corresponds to 15 µg of cytosolic extracts. c Left, Western blot of 20 and 40 µg of cytosolic extracts derived from siControl and of 13 and 26 µg of cytosolic extracts from siJMJD1B HeLa cells. These amounts of cytosolic proteins correspond to equal amounts of histone H3. Right, the graph shows the quantitation of global H3ac and H4K12ac, normalized by either the amount of histone H3 or H4. The data is showed as fold change with respect to siControl. The standard deviation was obtained from three independent experiments. *p < 0.05, Student’s t-test. d Left, Western blot of 20 µg of cytosolic extracts derived from Mock or overexpressed-Flag-tNASP HeLa cells. Right, the graph shows the quantitation of global H3ac and H4K12ac, normalized by either the amount of histone H3 or H4. The data is showed as fold change with respect to Mock. The standard deviation was obtained from three independent experiments. ***p < 0.001, Student’s t-test

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