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. 2018 Mar 7;8:4111. doi: 10.1038/s41598-018-22484-6

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© The Author(s) 2018

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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Inactivation of GADD45A impairs the viability and colony formation ability of cisplatin-treated melanoma cells. (A) Quantitative real-time PCR analyzing the mRNA expression of GADD45A in GADD45A-siRNA transfected A375 and SK-MEL-28 cells. (B) Western blot analyzed the protein expression of GADD45A in GADD45A-siRNA transfected A375 and SK-MEL-28 cells. (C) The protein expression of GADD45A in GADD45A-siRNA transfected and untransfected A375 cells following cisplatin treatment (4 μM, 48 h) by western blot. Protein expressions were normalized to the level of β-actin. (D) The cells were treated with cisplatin (0, 2, 4, 8 or 16 μM) for 48 h. The viability of the cells was detected by MTT assay and expressed relative to that of non-transfected cells. (E) The cells were treated with cisplatin (2 μM) for 48 h. After 15 days the surviving colonies were stained and counted. The colony numbers were counted, and the colony formation ratio was calculated according to the formula: Colony formation ratio = (number of colonies/number of cells seeded). Histogram shows the colony formation ratio (+SD) of GADD45A-siRNA transfected cells relative to control cells determined from three independent experiments. One-Way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001.