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. 2021 Feb 22;22(4):2172. doi: 10.3390/ijms22042172

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Interaction of EDEM3 mutants with NHK and soluble tyrosinase. (A) Analysis of ST association with EDEM3 constructs. Co-transfected lysed cells were divided in two: one half was immunoprecipitated with anti-tyrosinase antibodies while the other was immunoprecipitated with anti-HA antibodies. Immunoblots of isolated immunocomplexes were visualized with the indicated antibodies. TL indicates total lysates. (B) Analysis of NHK association with EDEM3 constructs. Cells were co-transfected with NHK and EDEM3 plasmids and processed as in (A), with the exception that anti-alpha-1 antitrypsin (A1AT) and anti-EDEM3 antibodies were used. TL indicates total lysates. (C) Analysis of MAN construct association with NHK. Due to the fact that commercial antibody used in (B) does not recognize MAN-EDEM3, an EDEM3 home-made antisera was used instead for immunoprecipitation after cell pulse labeling.