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. 2021 Sep 2;11:17587. doi: 10.1038/s41598-021-97060-6

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Interaction of CXXC5 and MeCP2. (a) The intracellular localization of CXXC5 and MeCP2 when co-synthesized is assessed with ICC of HEK293 cells transiently transfected for 48 h with the expression vector bearing the 3F-CXXC5 or the HA-MeCP2 cDNA. The Flag (green channel) or the HA (red channel) antibody was used to detect 3F-CXXC5 and HA-MeCP2, respectively. DAPI was used for DNA staining. The scale bar is 10 μm. (b,c) To examine the co-synthesis of CXXC5 and MeCP2, nuclear extracts of HEK293 cells transiently transfected with the expression vector bearing (b) the 3F-CXXC5 and/or the HA-MeCP2 or (c) HA-CXXC5 and/or the 3F-MeCP2 cDNA were subjected to WB analysis. Proteins were immunoblotted (IB) with the Flag or HA antibody. HDAC1 used as a loading control was probed with the HDAC1 antibody. (d,e) The nuclear extracts of HEK293 cells co-synthesizing 3F-CXXC5 and HA-MeCP2, or co-synthesizing HA-CXXC5 and 3F-MeCP2 were subjected to Co-IP using the HA antibody or the isotype-matched IgG followed by immunoblotting using (d) the Flag or (e) the HA antibody. Molecular masses (MM) in kDa are indicated.