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. 2018 Dec 5;9:5200. doi: 10.1038/s41467-018-07258-y

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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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Fig. 7 — Inhibition of CDK6 confers drug sensitivity to AKTi. a Left panel: MK2206-resistant cell lines were established by growing T47D and ZR75 cells in increasing concentrations of MK2206 for 4 months. Inductions of CDK6 expression was examined by western blot. Middle panel: the growth suppressive effect of MK2206 was presented by the percentage of S-phase decrease in parental or resistant cells after 2 μM MK2206 treatment for 4 days. Data are representative of three experiments performed in triplicate as the mean ± SD. ^*p < 0.01 when resistant cells were compared with parental cells. Right panel: the growth inhibition of these cells responded to the combination of MK2206 with JQ1 or Palbociclib was measured by cell count assays. Data are representative of three experiments performed in triplicate as the mean ± SD. ^*p < 0.01 when MK2206-treated group was compared with untreated group in parental cells; ^#p > 0.05 when MK2206-treated group was compared with untreated group in resistant cells. b Left panel: induction of CDK6 expression in parental and CDK6-overexpressing cells were examined by western blotting. Middle panel: the growth suppressive effect of MK2206 was presented by the percentage of S-phase decreased in parental or CDK6-overexpressing cells after 2 μM MK2206 treatment for 4 days. Data are representative of three experiments performed in triplicate as the mean ± SD. ^*p < 0.01 when CDK6-expressing cells were compared with vector control cells. Right panel: cell count assay was used to detect the growth inhibition effects of the combination of MK2206 with JQ1 or Palbociclib. Data are representative of three experiments performed in triplicate as the mean ± SD. ^**p < 0.01 when MK2206 plus JQ1 treatment group in CDK6-expressing cells was compared with corresponding group in parental cells. c Lapatinib (1 μM) also induced FOXO3a acetylation, its association with BRD4 and CDK6 expression in BT474 cells as analyzed by IP-western blotting. The growth inhibition effects of the combination of Lapatinib with JQ1 or Palbociclib on BT474 cells were reflected by cell count assays. Data are representative of three experiments performed in triplicate as the mean ± SD. ^*p < 0.01 when combinatory treatment group was compared with Lapatinib-treatment group