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. 2018 Nov 9;9:4728. doi: 10.1038/s41467-018-07188-9

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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. 5 — AMPK-Skp2-Akt axis is critical for survival under stress and glucose deprivation induced VEGF secretion and EGF-induced glycolysis and migration. a, c Cell survival analysis of MDA-MB-231 cells with control (shLuc), AMPKα1 knockdown, AMPKα1 knockdown along with Myr-Akt, Skp2 WT, or Skp2 S256A or Skp2 S256D restoration under hypoxia (1% O₂) for 72 h. b, d Cell survival analysis of MDA-MB-231 cells with control (shLuc), AMPKα1 knockdown, AMPKα1 knockdown along with Myr-Akt, Skp2 WT, or Skp2 S256A or Skp2 S256D restoration under glucose deprivation for 16 h. e MDA-MB-231 cells with control (shLuc), AMPKα1 knockdown and AMPKα1 knockdown with vector control, Skp2 WT and S256A or Skp2 S256D restoration were treated with a glucose-free medium for 8 h, and supernatant was collected for HUVEC tube formation assay. Each value represents the mean ± SEM in three independent experiments. **P < 0.01. f MDA-MB-231 cells with control (shLuc), AMPKα1 knockdown (#1) and Myr-Akt restoration in AMPKα1 knockdown were starved in DMEM glucose-free medium for 4 h and added with 2-NBDG for 30 min. Cells were then subjected to FACS analysis. Each value represents the mean ± SEM (n = 4 per group) in three independent experiments. **P < 0.01. g Immunoblotting of control (shLuc) and AMPKα knockdown (#1) MDA-MB-231 cells serum starved and treated with EGF for 15 and 30 min. h Immunoblotting for MDA-MB-231 cells with control (shLuc), AMPKα1 knockdown and AMPKα1 knockdown along with Myr-Akt restoration. i MDA-MB-231 AMPKα1 knockdown cells with Skp2 WT, S256A or S256D restoration were starved in DMEM glucose-free medium for 4 h and added with 2-NBDG for 30 min. Cells were then subjected to FACS analysis. Each value represents the mean ± SEM (n = 4 per group) in three independent experiments. **P < 0.01. j Cell migration assay of MDA-MB-231 cells with control (shLuc), AMPKα1 knockdown and AMPKα1 knockdown along with Myr-Akt restoration. k MDA-MB-231 AMPKα1 knockdown cells with Skp2 WT, S256A or S256D restoration were subject to cell migration assay with or without EGF