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. 2020 Jun 8;11(6):434. doi: 10.1038/s41419-020-2633-7

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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 The relative expression of FOXC2-AS1 in the cytoplasm and nucleus of SW480 and LoVo cells. FISH detection was used to investigate the location of FOXC2-AS1 in cells (b) and tissues (c). Scale bars = 100 μm. d The expression of FOXC2 was examined in 66 CRC and 15 adjacent normal tissues. e The expression relationship between FOXC2-AS1 and FOXC2 was analyzed in 66 CRC tissues. FOXC2 expression was examined in FOXC2-AS1-silenced SW480 and LoVo cells by qRT-PCR method (f) and Western blot (g). h qRT-PCR detected the levels of nascent FOXC2 pre-mRNA with Click-iT Nascent RNA Capture Kit in FOXC2-AS1-silenced and control cells. i qRT-PCR investigated FOXC2 mRNA stability in FOXC2-AS1-silenced and control cells treated with the transcriptional inhibitor actinomycin D (50 ng/ml) for different times. j Schematic diagram of FOXC2-AS1 and FOXC2 gene locus and structure. The red grid represents the completely complementary region. The number represents the length of exon or intron. k qRT-PCR was conducted to analyze RNase protection experiment. β-actin was used as a negative control, while PDCD4-AS1 was used as a positive control. l qRT-PCR was performed to assess the interaction between FOXC2 and biotin-labeled FOXC2-AS1 after RNA pull-down assay. *P < 0.05, ***P < 0.001, N.S. indicates not statistically different.