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. 2021 Jul 7;49(14):8261–8276. doi: 10.1093/nar/gkab579

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 7. — Inhibition of polysome association of CTIF by DDX19B mainly occurs on CBC-bound mRNA. HEK293FT cells stably expressing Myc-eIF4E were transiently co-transfected with a plasmid expressing CBP80-HA and a plasmid expressing either FLAG-CTIF-WT-KASH1 or FLAG-CTIF-F460A-KASH1. The cytoplasmic extracts were subjected to polysome fractionation. Next, total fractions were pooled into seven fractions and subjected to IP with either α-HA antibody or α-Myc antibody. (A) Relative distribution of the indicated proteins. Protein samples in each fraction before or after IP were analyzed by western blotting. (B and C) Quantitative representation of the relative distribution of FLAG-CTIF-WT-KASH1 and FLAG-CTIF-F460A-KASH1 across the gradient before (B) and after IP (C), as shown in Figure 5B–D; n = 3.