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. 2018 Oct 31;9:4541. doi: 10.1038/s41467-018-06808-8

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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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Fig. 5 — eIF4E3 and HDAC1 promote MB growth and inhibit VCR sensitivity of MB cells. a Proliferation of D556Med cell transfected with scrambled- or eIF4E3/HDAC1-siRNA. Cell proliferation was measured using an IncuCyte phase-only processing module. The p-values were determined by the sum-of-squares F test. Error bars represent mean ± SEM of three independent experiments. b Difference in morphology between D556Med cells transfected with scrambled-siRNA, eIF4E3-siRNA, or HDAC1-siRNA. c Colony-forming assay of D556Med cells transfected with scrambled-siRNA, eIF4E3-siRNA, or HDAC1-siRNA. Bar graph shows crystal violet-stained colonies counted microscopically. The p-value was calculated using a standard Student t-test. Error bars represent mean ± SEM of three independent experiments (performed in triplicate/experiment). d Photomicrograph showing migrated D556Med cells transfected with scrambled-siRNA, eIF4E3-siRNA, or HDAC1-siRNA. Bar graph shows number of migrated cells microscopically counted in ten different fields. The p-value was calculated using a standard Student t-test. Error bars represent mean ± SEM of three independent experiments. e Line graphs show mean tumor volume in nude mice intracranially injected with DAOY–GFP–luciferase cells transfected with scrambled-siRNA or eIF4E3/HDAC1-siRNA (n = 6 mice/group). The p-values were determined by the sum-of-squares F test. Error bars represent mean ± SEM. f Live bioluminescence (BLI) images of mice injected with DAOY–GFP–luciferase cells transfected with scrambled-siRNA or eIF4E3/HDAC1-siRNA (n = 6 mice/group). g VCR dose–response curve of D556Med cells transfected with scrambled-siRNA, eIF4E3-siRNA or HDAC1-siRNA and treated with VCR or vehicle. Cell viability was assessed using alamarBlue cell viability assay. The p-values were determined by the sum-of-squares F test. Error bars represent mean ± SEM of three independent experiments (performed in sixtuplicate/experiment). h miR-584-5p rescues cancer growth-promoting effects of eIF4E3 and HDAC1. Bar graphs showing viability of D556Med cells transfected with control plasmid or eIF4E3/HDAC1 expression vector or cotransfected with eIF4E3/HDAC1 expression vector and miR-584-5p mimic. The p-value was calculated using one-way analysis of variance (ANOVA) followed by Sidak’s multiple-comparisons test. Error bars represent mean ± SEM of three independent experiments (performed in sixtuplicate/experiment). ****p < 0.0001; ***p < 0.001; **p < 0.01. Scale bars: 50 μM (b, d)