Prolonged S-phase of cell cycle in eIF5B-knockdown cells.

<p>(A) Quantitative PCR analysis of mRNA expression levels of DNA polymerases in the DNA synthesis pathway; (B) Nucleotides accumulated within cells; and (C) Cell numbers in each cell cycle phase as determined by flow cytometry (*<i>p</i><0.05; **<i>p</i><0.01; ***<i>p</i><0.001; n = 3).</p

Quantitative PCR and western blot analysis of the eIF5B-knockdown induced deactivation of MAPK signaling pathways.

<p>(A) Quantitative PCR analysis of mRNA expression of selected genes in MAPK signaling pathways (*<i>p</i><0.05; **<i>p</i><0.01; ***<i>p</i><0.001; n = 3); and (B) Total cell lysates immunoblotted with MEK1, Erk1/2, phospho-Erk1/2 (Thr202/Tyr204), MEK3, p38 and phospho-p38 (Thr180/Tyr182) antibodies.</p

Amino acid accumulation and autophagy induced in eIF5B knockdown cells.

<p>(A) Quantitative PCR analysis of mRNA expression of essential amino acid transporters; (B) Levels of intracellular amino acids in eIF5B knockdown cells compared with control cells; (C) Protein expression levels of LC3 and p62 following cell culture for 2 h in the presence or absence of 100 nM Baf-A1, as determined by western blotting; (D) Total cell lysates immunoblotted with mTOR, phospho-mTOR (Ser2448), P70S6K and phospho-P70S6K (Thr389) antibodies; (E) RT-PCR analysis of rRNA concentrations using rRNA-specific primers listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168387#pone.0168387.s010" target="_blank">S1 Table</a> (*<i>p</i><0.05; **<i>p</i><0.01; ***<i>p</i><0.001; n = 3).</p

Functional classification of proteins downregulated in eIF5B knockdown cells.

<p>Functional classification of proteins downregulated in eIF5B knockdown cells.</p

Characterization of eIF5B-KN1-293T and control cells.

<p>(A) Semi-quantitative RT-PCR analysis of eIF5B mRNA levels; (B) Western blot analysis of eIF5B protein expression; (C) Cellular ROS levels; (D) Survival rates of cells treated with different concentrations of H₂O₂ for 12 h, as determined by trypan blue dye exclusion assay; and (E) Cell growth curves. Data are presented as the mean and standard deviation (*<i>p</i><0.05; **<i>p</i><0.01; ***<i>p</i><0.001; n = 3).</p

Additional file 1: Figure S1. of Kinetic and thermodynamic studies reveal chemokine homologues CC11 and CC24 with an almost identical tertiary structure have different folding pathways

Comparison of folding and unfolding trances of CCL11 and CCL24 fitted with different exponentials. (A) Kinetics traces of CCL11 at 1.5 M GdnHCl fitted with monophasic relaxation eq. (B) Kinetics traces of CCL11 at 1.5 M GdnHCl fitted with biphasic relaxation eq. (C) Kinetics traces of CCL11 at 6 M GdnHCl fitted with monophasic relaxation eq. (D) Kinetics traces of CCL11 at 6 M GdnHCl fitted with biphasic relaxation eq. (E) Kinetics traces of CCL24 at 4 M GdnHCl fitted with monophasic relaxation eq. (F) Kinetics traces of CCL24 at 4 M GdnHCl fitted with biphasic relaxation eq. (G) Kinetics traces of CCL24 at 6 M GdnHCl fitted with monophasic relaxation eq. (H) Kinetics traces of CCL24 at 6 M GdnHCl fitted with biphasic relaxation equation. (a-h) represent the residues of the fits corresponding to (A-H) respectively. (DOCX 1613 kb