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. 2018 May 10;8:7459. doi: 10.1038/s41598-018-25798-7

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Knockdown of eIF4G1 impairs mRNA translation and sensitizes CRPC cells to current therapy and eIF4G complex inhibitor impairs colony formation in CRPC cells: Cytoplasmic extracts from siControl and sieIF4G1 for LNCaP (a) & C4-2B (b) cells were centrifuged respectively through continuous sucrose gradients to obtain fractions of increasing density. Absorbance (A₂₅₄) (y-axis) versus increasing density (x-axis) of fractions was monitored for the presence of ribosomal subunits, ribosomes, and polysome. Polysome (P)/Monosome (M) Ratio was calculated further. (c) Western blotting showing knockdown of eIF4G1 in LNCaP and C4-2B. (d) The graph is quantitation of western blot for eIF4G1. (e) Representative image of cell viability assay done by crystal violet with CRPC (C4-2B cell line) cells with siControl and sieIF4G1 treated with 10 µM Enzalutamide (ENZ) and 10 µM Bicalutamide (BIC) for 24 h. (f) The Graph showed quantitation for above plate absorbance at 560 nm. (g) Colony formation assay for CRPC cells (C4-2B) with 10 µM ENZ, 10 µM 4EGI-1 (eIF4G-eIF4E complex inhibitor) and combination of 10 µM 4EGI-1 and 10 µM ENZ treatment. (h) The graph is the number of colonies counted. Colonies were counted using an ImageJ software. Error bars represent ± SD. p-values are indicated as *<0.05, **<0.01, ***<0.001.