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. 2021 Jun 18;12:3734. doi: 10.1038/s41467-021-24099-4

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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 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 Representative microphotographs of in situ hybridization (ISH) analysis of PLANE expression. DapB: negative control; PPIB: positive control. Scale bar, 10 µm. Data are representatives of 3 independent experiments. b qPCR analysis showing PLANE subcellular localization. Cyt: cytoplasm; Nuc: nucleus. β-actin: Cyt marker; U6: Nuc marker. Data are mean ± s.d.; n = 3 independent experiments, two-tailed Student’s t-test. c In vitro-synthesized PLANE bound to the PLANE-binding region (PLANE-BR) of in vitro-transcribed intron 45 of NCOR2 pre-mRNA as shown in domain-specific chromatin isolation by RNA purification (dChIRP) assays. This binding was abolished when the PLANE-BR at NCOR2 pre-mRNA or the duplex-forming oligonucleotides (DFO) within PLANE were deleted (intron 45-∆PLANE-BR and PLANE-∆DFO, respectively). Data are representatives of three independent experiments. d In vitro-transcribed PLANE did not precipitate in vitro-transcribed intron 47 of the NCOR2 pre-mRNA that does not contain the PLANE-BR as shown in dChIRP assays. Data are representatives of 3 independent experiments. e In vitro-synthesized PLANE precipitated the endogenous NCOR2 pre-mRNA but not NCOR2 genomic DNA as shown in dChIRP assays. Data are representatives of three independent experiments. f Endogenous PLANE co-precipitated a PLANE-BR-containing fragment but not a non-PLANE-BR-containing fragment of intron 47 of NCOR2 pre-mRNA as shown in dChIRP assays. Data are representatives of three independent experiments. S, sense; AS, antisense. g PLANE co-precipitated a fragment of intron 45 of the NCOR2 pre-mRNA in A549 cells treated with proteinase K as shown in dChIRP assays. Data are representatives of three independent experiments. h, i PLANE-∆DFO did not affect the NCOR2-202-generating AS event (h) and 5-bromo-2′-deoxyuridine (BrdU) incorporation (i). Data are representatives or mean ± s.d.; n = 3 independent experiments, two-tailed Student’s t-test. j, k A shRNA-resistant PLANE (PLANE-R) diminished the enhancement of the NCOR2-202-generating AS event caused by induced PLANE knockdown as shown in RT-PCR (j) and the relative quantification using densitometry (k). Data are representatives or mean ± s.d.; n = three independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. Source data are provided as a Source data file.