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. 2022 Aug 17;15:112. doi: 10.1186/s13045-022-01338-9

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — LINC00623 blocks NAT10 ubiquitination and degradation. a The protein levels of NAT10 in BxPC-3 and PANC-1 cells. β-Actin was used as the loading control. b The protein levels of NAT10 in the indicated cells treated with cycloheximide (CHX, 50 μg/mL) for 0, 2, 4, 6, 8, and 10 h. c The protein levels of NAT10 in the indicated cells treated with or without MG132 (10 μM) for 6 h. d BxPC-3 cell lysates were immunoprecipitated (IP) with an anti-NAT10 antibody and immunoblotted with an anti-HA antibody. e IP was performed to detect the association between NAT10 and USP39 in BxPC-3 cells. d–e BxPC-3 cells were transiently transfected with LINC00623 knockdown, LINC00623 overexpression or control plasmids and then treated with MG132 for 4 h before cell lysates were harvested for co-IP with an anti-NAT10 (IP: NAT10) or anti-USP39 (IP: USP39) antibody. f The colocalization of NAT10 (green) and USP39 (red) in BxPC-3 cells was evaluated by IF. Scale bar = 2 μm (left). The percentage of colocalization (Mander’s coefficients) of NAT10 and USP39 is summarized in the bar charts (right). g The ubiquitination levels of NAT10 were determined by Western blotting. BxPC-3 cells were transiently transfected with USP39 silencing (left), USP39 overexpression (right) and control plasmids and then treated with MG132 for 4 h before cell lysates were harvested for co-IP with an anti-NAT10 antibody and immunoblotted with an anti-HA antibody. h The ubiquitination levels of NAT10 in the products of IP with an anti-Flag antibody in lysates from BxPC-3 cells transfected with LINC00623 knockdown or USP39 overexpression and control plasmids