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. 2020 Apr 3;10:448. doi: 10.3389/fonc.2020.00448

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Copyright © 2020 Felce, Anderson, Maguire, Gascoyne, Armstrong, Wong, Li and Banham.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of A20 CRISPR/Cas9 Foxp1 knockout clones. (A) Exon coding DNA from selected clones (E6.1: A20 E6 clone 1; E6.2: A20 E6 clone 2; E7.1: A20 E7 clone 1; E7.2: A20 E7 clone 2) was analyzed by Sanger sequencing and compared to wild-type (WT) Foxp1. Each clone contained a deletion or insertion leading to a frameshift and the introduction of a premature stop codon. (B) Western blotting of biological replicates, used for RNA sequencing analysis, showed that knockout of Foxp1 protein expression was stable during subsequent passages of each clone. (C) MHC-II expression was elevated in all of the clones, while Cd74 expression was unaltered or inconsistently reduced, compared to A20 parental cells.