Fig. 1

siJMJD1B cells accumulate tNASP, Hsp90, H3, and H4. a Western blot of 10 and 20 µg of cytosolic and nuclear extracts derived from HeLa cells. As control of the extract preparation, HIRA and Topo-I were analyzed as nuclear extract markers, showing that cytosolic extract is not contaminated with nuclear components. TE, total cell extract derived from HeLa cells. b Western blot of 5 and 15 µg of cytosolic extracts derived from either siControl or siJMJD1B treated HeLa cells. c Levels of mRNA from samples derived from siControl or siJMJD1B treated HeLa cells measured by real-time PCR. The graph shows the fold change of the different mRNAs relative to siControl. Each expression level was normalized to that of GAPDH. The standard deviation was obtained from three independent experiments. ***p < 0.001, Student’s t-test. d Cells were treated with either siControl or siJMJD1B for 48 h. Then, cells were treated with either empty or the JMJD1B derived JmjC-domain containing vector, for 24 h. Cytosolic extracts were prepared and analyzed by Western blot. To the left, Western blot of 20 µg of cytosolic extracts derived from either siControl or siJMJD1B treated HeLa cells that have been transcomplemented with either empty vector or JmjC-domain. To the right, graphs showing the quantitation of tNASP, sNASP, and histone H3 normalized by β-actin. The standard deviation was obtained from three independent experiments. ***p < 0.001, Student’s t-test