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. 2019 Jul 17;9:10369. doi: 10.1038/s41598-019-46594-x

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© The Author(s) 2019

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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Elevated DNFA expression accompanies drug resistance in BRAFi treated melanoma cells. (a) RT-qPCR assay compared the DNFA gene expression between HT-144 cells in 10% FBS medium and HT-144BR cells in 10% FBS medium with vemurafenib (2 μM). (b) RT-qPCR assay compared the DNFA gene expression between LOXIMVI cells in 10% FBS medium and LOXIMVIBR cells in 10% FBS medium with vemurafenib (2 μM). (c,d) HT-144 cells were treated with different dosages of vemurafenib for 0.5 or 1 day in 1% ITS medium. SCD expression was assayed by RT-qPCR analysis. Expression of SCD from all treatment groups was normalized to expression under DMSO treatment at day 0.5 (normalized as 1). Relative gene expression was compared between 0.5-day and 1-day treatment groups. Data were presented as mean ± SD and quantified from triplicates. Two-way ANOVA tests were performed. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. (e–g) HT-144 cells were transfected with ASO-4 (5 nM) and then treated with vemurafenib (100 nM) in 1% ITS medium for one day. RT-qPCR assay analyzed DNFA gene expression from treatment groups relative to expression under ASO-Neg and DMSO treatment at day 1 (normalized as 1). Data are presented as mean ± SD and quantified from triplicates. One-way ANOVA tests were performed. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. (h) HT-144 cells were transfected with ASO-4, and then treated in combination with vemurafenib. Cell viability was measured by cell titer-glo assay three days after combined treatment in 1% ITS medium. (i) The drug interaction in (h) was evaluated with the Bliss independence model.