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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. 3 — LINC00623 binds to NAT10. a GO-CC (cellular component) analysis predicting the potential proteins binding to LINC00623. b Western blotting was performed to evaluate the specific association of NAT10 with biotinylated LINC00623. Lysates of BxPC-3 and PANC-1 cells were harvested for RNA pulldown assays. LINC00623 antisense RNA was used as the negative control. c A RNA-binding protein immunoprecipitation (RIP) assay was performed using an antibody against NAT10. RT-PCR was used to detect LINC00623 enrichment. IgG was used as the isotype control. d Immunofluorescence (IF) staining showed that LINC00623 (red) was colocalized with NAT10 (green) in the nucleus. Scale bar = 10 μm. e Western blotting of NAT10 in samples precipitated by biotinylated full-length LINC00623 (FL) or LINC00623 truncations (Δ1: 1–699 bp; Δ2: 700–1200 bp; Δ3: 1201–1800 bp; Δ4: 1801–2500 bp; Δ5: 2501–3000 bp; Δ6: 3001–3607 bp). FL LINC00623 was used as the positive control. f Flag-RIP assay for LINC00623 showing its fold enrichment in cells transiently transfected with plasmids containing FLAG-tagged full-length and truncated NAT10 constructs (Del1: 1–280 aa; Del2: 281–488 aa; Del3: 558–753 aa; Del4: 753–1052 aa). IgG-RIP was used as the internal control. The values are expressed as the mean ± SD of three independent experiments