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. 2020 Apr 23;11(4):265. doi: 10.1038/s41419-020-2432-1

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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. 2 — a Western blotting analysis of cell-cycle-related regulators in RAGE-overexpressed subclones of A549 was shown; b p21^CIP1 mRNA expression was measured by quantitative real-time PCR in A549 cells after transient transfection of RAGE; c A549 subclones were transfected with control siRNA or siRNA against CDKN1A (p21^CIP1). Cell growth was measured to evaluate the effect of p21^CIP1 knockdown in RAGE-overexpressed A549 subclones; d protein interactions between p21^CIP1-CDK2/4 and RB-CDK2/4 were evaluated by immunoprecipitation; e the kinase activities of CDK2 or CDK4 were detected by in vitro kinase assay while Histone H1 was used as the specific substrate; f the levels of CDK2-specific T821 residue phosphorylation of RB(P-RB^T821) and CDK4-specific S780 residue (P-RBS⁷⁸⁰) were detected by western blotting analysis; g p53 was knockdown by transfection of siRNA in A549 subclones and the protein levels of p21^CIP1 and p53 were measured. β-actin served as the internal control. P stands for parental and R^low/R^high stands for different clones of the cells. *p < 0.05; **p < 0.01 compared to parental cells.