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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 6. — DDX19B inhibits polysome association of CTIF in the perinuclear region. (A) Schematic illustration of FLAG-CTIF-WT-KASH1 and FLAG-CTIF-F460A-KASH1. The FLAG tag, CBP80-interacting domain, MIF4G domain, and KASH1 domain are depicted. The position of the F460A mutation is indicated by a black arrow. (B) Immunostaining of FLAG-CTIF-WT and its variants. HeLa cells transiently expressing the indicated protein were subjected to immunostaining with α-FLAG antibody (green) and DAPI (blue); n = 2; Scale bar, 10 μm. (C) Polysome fractionation analysis of cytoplasmic extracts of HEK293FT cells transiently expressing FLAG-CTIF-WT-KASH1 or FLAG-CTIF-F460A-KASH1. The protein samples in each fraction after polysome fractionation experiments were analyzed by western blotting with α-FLAG antibody. (D) Quantitative representation of relative distributions of FLAG-CTIF-WT-KASH1 or FLAG-CTIF-F460A-KASH1. Mean values and standard deviations from three biological replicates are shown; n = 3; #, not significant; *P < 0.05, **P < 0.01.