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. 2019 Sep 10;9:12973. doi: 10.1038/s41598-019-49480-8

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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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Anoikis partly contributes to the in vivo effect of the combination of knockdown of PDK1 and CPT. (A) HCT116 cells with or without the transduction of a PDK1 shRNA were seeded into 6-well plates and cultured in normal conditions. Then the cells were stained with AnnexinV-FITC/PI. Flow cytometry showed that the apoptosis rate was much higher after PDK1 was silenced. (B) As indicated in (A), the cells were seeded into 6-well plates coated with poly-HEMA. Flow cytometry showed that the apoptotic rate of HCT116 cells stably transduced with PDK1 was significantly higher than the other two controls. (C) Histogram indicated the corresponding comparison of the apoptosis rate presented in (A,B). (D) The results showed knockdown of PDK1 strongly sensitized CRC to anoikis; however, the anoikis rate was significantly reduced when PDK1 silencing was combined with CPT. (E) Histogram indicated the corresponding comparison of the apoptosis results presented in both normal condition and suspending condition in (D). Data expressed as mean ± S.D., *** represents P < 0.001, and **** represents P < 0.0001.