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. 2021 Apr 30;12(5):430. doi: 10.1038/s41419-021-03721-9

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© The Author(s) 2021, corrected publication 2024

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 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/.

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Fig. 4 — A, B H1299 cells were transiently transfected with control or GRHL1 plasmids. Forty-eight hours later, total RNAs were extracted. The mRNA levels were determined by q-PCR. Data represent the average of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ns: P > 0.05. C H1299 cells were transiently transfected with control or GRHL1 siRNAs. Forty-eight hours later, total RNAs were extracted. The mRNA levels were determined by q-PCR. Data represent the average of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. D H292 cells were transiently transfected with control or His-GRHL1 plasmids. Forty-eight hours later, the cells were lysed. Protein expression was assessed by western blotting using the indicated antibodies. E H1299 cells were transiently transfected with control or GRHL1 siRNAs. Forty-eight hours later, the cells were lysed. Protein expression was assessed by western blotting using the indicated antibodies. F In 293 T cells, ChIP assay was conducted with anti-His and control rabbit IgG for immunoprecipitation, followed by PCR with CDC27, RAD21, CDC7, and ANAPC13 promoter-specific primers. G pGL3-enhancer vector containing CDC27, RAD21, CDC7, and ANAPC13 promoter fragment was transfected into H1299 respectively, co-transfected with Renilla control plasmid and pcDNA3.1 vector or pcDNA3.1-GRHL1 plasmid. The relative levels of luciferase activity were normalized to the levels of vector control and to the levels of luciferase activity of the Renilla control plasmid. Data represent the average of three independent experiments (mean ± SD). *P < 0.05, **P < 0.01. H pGL3-enhancer vector containing CDC27, RAD21, CDC7, and ANAPC13 promoter fragment was transfected into H1299 respectively, co-transfected with Renilla control plasmid and control siRNA or GRHL1 siRNAs. The relative levels of luciferase activity were normalized to the levels of vector control and the levels of luciferase activity of the Renilla control plasmid. Data represent the average of three independent experiments (mean ± SD). *P < 0.05. I pGL3-enhancer vector containing CDC27-mutation, RAD21-mutation, CDC7-mutation, and ANAPC13-mutation promoter fragment was transfected into H1299 respectively, co-transfected with Renilla control plasmid and pcDNA3.1 vector or pcDNA3.1-GRHL1 plasmid. The relative levels of luciferase activity were normalized to the levels of vector control and the levels of luciferase activity of the Renilla control plasmid. Data represent the average of three independent experiments (mean ± SD). ns P > 0.05.