KDM4A levels affect the distribution of translation initiation factors

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KDM4A levels affect the distribution of translation initiation factors. KDM4A levels affect the distribution of translation initiation factors. A and B, fractions collected from the polysome profiles [Negative Control (Negative) or KDM4A depleted HEK 293T cells for 48 hours (A); HEK 293T cells overexpressing WT or H188A for 24 hours (B)] were immunoblotted with the indicated antibodies. Representative Western blot analyses are shown below the graphs and represent the average of the quantification by ImageJ from two independent experiments. The red bars represent a reproducible increase of at least 20% compared with the control fractions from independent KDM4A siRNA–treated cells or KDM4A WT- or H188A-overexpressing cells (see individual experiments in Supplementary Fig. S1C and S1D). The top Western blot analysis represents whole cell extracts or the input extracts used to generate the polysome profiles. C, fractions collected from the polysome profiles from HEK 293T cells overexpressing GFP, WT, or H188A for 24 hours were immunoblotted with anti-GFP and the corresponding graph represents the average from two independent experiments. The y-axis represents the fold change for each fraction relative to the fraction five from the corresponding control. For A, the negative siRNA is the control, whereas for B, the GFP is the control. For C, the GFP-WT is the control. The red bars represent a reproducible increase of at least 20% compared with the GFP-WT fractions between two independent experiments. D, schematic representing where KDM4A could directly be involved in translation. Also see Supplementary Fig. S1. Capucine Van Rechem et al. Cancer Discovery 2015;5:255-263 ©2015 by American Association for Cancer Research