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. 2019 Oct 28;17:137. doi: 10.1186/s12964-019-0446-z

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 5 — Colony formation assay in HCC827 cell lines and western blotting in 3 cell lines by mono therapy or combination treatment of OTSSP167 plus auranofin. The combination of OTSSP167 and auranofin potentiates colony formation inhibition in HCC827 cell lines. Combination of OTSSP167 and auranofin downregulates expression and inhibits activation of PAK1 and PKCι. a HCC827 cell colonies were grown under single or combination treatment (OTSSP167 and auranofin) at indicated dose. Fixed colonies were stained using crystal violet. b Concentration of crystal violet was represented as ratio to control (non-treatment colonies defined as 1). Crystal violet was absorbed using 2% sodium dodecyl sulfate and measured at 570 nm. c Combination index by each concentration of OTSSP167 and auranofin. Combination index decreased in dose-dependent manner. Combination index was included in synergistic area (< 0.9). d HCC827, H23 and H520 cell lines were exposed to 3 or 10 μM auranofin (10 μM for HCC827, 3 μM for H23 and H520) or 1 μM OTSSP167 and the combination of OTSSP167 plus auranofin for 6 h. Protein expression and activation were analyzed by western blotting. Actin was used as house-keeping protein. The experiments were made at least twice. The combination of OTSSP167 plus auranofin abrogated the expression and phosphorylation of PAK1 and PKCι. Abbreviations: S, serine; T, threonine; Y, tyrosine