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. 2018 Dec 13;19(12):4032. doi: 10.3390/ijms19124032

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Depletion of eIF5B leads to increased translation of ATF4. HEK293T cells were reverse-transfected with a non-specific control siRNA (siC) or an eIF5B-specific siRNA pool (si5B), incubated 96 h, harvested in RNA lysis buffer, and subjected to polysome profiling analysis. (A) A representative polysome profile from control versus eIF5B-depleted HEK293T cells. (B) The proportion of ATF4 mRNA (relative to β-Actin mRNA) for each fraction from the panel (A). (C) Fractions 1–2 (representing monosomes) were pooled, as were fractions 3–8 (representing polysomes). The ratio of polysomes/monosomes is shown for a representative experiment. Data in panels (B,C) are expressed as the mean ± SD for technical triplicates. An independent experiment is shown in Figure S2.