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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. 2 — a–c SiRNA knockdown of PLANE (a) inhibited 5-bromo-2′-deoxyuridine (BrdU) incorporation (b) and clonogenicity (c) in multiple cancer cell lines. Relative clonogenicity was quantitated using ImageJ-plugin ‘ColonyArea’. Data are mean ± s.d. or representatives; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison test. Scale bar, 1 cm. d Induced knockdown of PLANE by the addition of doxycycline (Dox, 500 nM) in A549.shPLANE and H1299.shPLANE cells. Data are mean ± s.d.; n = 3 independent experiments, two-tailed Student’s t-test. e Dox (500 nM)-induced knockdown of PLANE inhibited A549.shPLANE and H1299.shPLANE cell proliferation as shown by decelerated cell number increases. Data are mean ± s.d.; n = 3 independent experiments, two-tailed Student’s t-test. f Induced knockdown of PLANE inhibited A549.shPLANE and H1299.shPLANE cell clonogenicity, which was partially reversed by cession of Dox (500 nM) treatment. Relative clonogenicity of cells was quantitated using ImageJ-plugin ‘ColonyArea’. Data are representatives or mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bar, 1 cm. g Representative microscopic photographs of anchorage-independent growth of A549.shPLANE and H1299.shPLANE cells with or without treatment with Dox and cessation of Dox treatment. Quantification of anchorage-independent growth of the cells is also shown. Scale bar, 0.5 mm. Data are representatives or mean ± s.d.; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparisons test. h, i Representative photographs (h) and growth curves (i) of A549.shPLANE xenografts in nu/nu mice with or without treatment with Dox (2 mg/ml supplemented with 10 mg/ml sucrose in drinking water) and cessation of Dox treatment. Data are representatives or mean ± s.d.; n = 6 mice per group, one-way ANOVA followed by Tukey’s multiple comparison test. DOX: 2 mg/ml supplemented with 10 mg/ml sucrose in drinking water. Source data are provided as a Source data file.