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. 2017 Nov 8;7:15031. doi: 10.1038/s41598-017-15172-4

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© The Author(s) 2017

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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Depletions of the FA pathway factors increased the sensitivity of gemcitabine to SK-MES-1 cells. (A) A549, SK-MES-1, KLN205, HCC4006 and Calu-1 cell lines growing in 96-well plates were treated with gemcitabine at the indicated dose for 4 h. The CCK-8 assay was used to determine cell viability (^*SK-MES-1 or Calu-1 as compared with A549, KLN205, and HCC4006 respectively, P < 0.05). (B) Western blot of FANCL-, FANCD2-, BRCA2- and CHK1-depleted SK-MES-1 cells to verify the efficiency of the siRNA transfections. β-actin was used as loading control. Uncropped images are presented in Supplementary Figure S5. (C) SK-MES-1 and (D) KLN205 cells growing in 96-well plates were transfected with various siRNAs as indicated. Cell viability was detected by the CCK-8 assay following gemcitabine treatment for 4 h (^*siContrl as compared with siFANCL and siFANCD2, P < 0.005, compared with siCHK1, P < 0.001; ^#siControl as compared with siFANCL and siFANCD2, P < 0.05, compared with siCHK1, P < 0.005). (E) SK-MES-1 and (F) KLN205 cells growing 96-well plates were transfected with various siRNA as indicated. Cell survival was measured by the CCK-8 assay following cisplatin treatment (siControl as compared with siFANCL and siFANCD2, *P < 0.01).