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. 2020 Aug 12;11(8):683. doi: 10.1038/s41419-020-02848-5

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© The Author(s) 2020

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

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Fig. 4 — a, b 293T cells were co-transfected with HA-tagged PAX6 and Flag-tagged SUMO1, as indicated. Whole-cell lysates were prepared 24 h post transfection under denaturing conditions and immunoprecipitated with anti-HA antibody (a) or anti-Flag antibody (b). Western blot analysis was used to detect SUMOylation-modified PAX6. c, d 293T cells were co-transfected with HA-tagged PAX6, Flag-tagged SUMO1, RGS-tagged SENP1, or RGS-tagged SENP1m as indicated. Whole-cell lysates were prepared 24 h post transfection under denaturing conditions and immunoprecipitated with anti-HA antibody (c) or anti-Flag antibody (d). Western blot analysis was used to detect de-SUMOylation of PAX6. e 293T cells were co-transfected with Flag-tagged PAX6, HA-tagged SUMO2/3, Myc-tagged Ubc9, or RGS-tagged SENP1 as indicated. Whole-cell lysates were prepared 24 h post transfection under denaturing conditions and immunoprecipitated with anti-Flag antibody. f Cell lysates of B4G12 were prepared under denaturing conditions and subjected to immunoprecipitation with anti-PAX6 antibody, followed by western blot using anti-PAX6, anti-SUMO1, or anti-GAPDH antibodies. The result of IP–WB showed the endogenous SUMOylation of PAX6.