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. 2020 Jul 13;36:101643. doi: 10.1016/j.redox.2020.101643

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© 2020 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 4 — Glutamine metabolism-mediated cancer cell survival is regulated by AMPK-Nrf2 under ECM detachment. (A, B, D, E) HepG2 cells expressing sh-con or sh-AMPK were cultured in attachment (AT) or poly-HEMA-coated (DT) dishes for 24 h. Western blot analysis was performed on protein extracts of these cells with antibodies against the indicated proteins, with either β-actin or lamin B1 as loading controls. (A, D) The asterisk indicates a nonspecific band. The expression levels of protein were measured by densitometric analysis. (C) Cells stably expressing sh-con were transfected with an empty vector. Cells stably expressing sh-AMPK were transfected with either an empty vector or the EGFP-Nrf2 expression vector. Western blot analysis was performed on protein extracts of these cell lines with antibodies against the indicated proteins after culturing in adherent or poly-HEMA-coated dishes for 24 h. Representative image of western blots are shown. (F) ROS levels in HepG2 cells stably expressing sh-con, sh-AMPK, or sh-Nrf2 for 24 h after plating in adherent or poly-HEMA-coated pates for 24 h. Data are presented as mean ± SD of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.

Fig. 4 — Glutamine metabolism-mediated cancer cell survival is regulated by AMPK-Nrf2 under ECM detachment. (A, B, D, E) HepG2 cells expressing sh-con or sh-AMPK were cultured in attachment (AT) or poly-HEMA-coated (DT) dishes for 24 h. Western blot analysis was performed on protein extracts of these cells with antibodies against the indicated proteins, with either β-actin or lamin B1 as loading controls. (A, D) The asterisk indicates a nonspecific band. The expression levels of protein were measured by densitometric analysis. (C) Cells stably expressing sh-con were transfected with an empty vector. Cells stably expressing sh-AMPK were transfected with either an empty vector or the EGFP-Nrf2 expression vector. Western blot analysis was performed on protein extracts of these cell lines with antibodies against the indicated proteins after culturing in adherent or poly-HEMA-coated dishes for 24 h. Representative image of western blots are shown. (F) ROS levels in HepG2 cells stably expressing sh-con, sh-AMPK, or sh-Nrf2 for 24 h after plating in adherent or poly-HEMA-coated pates for 24 h. Data are presented as mean ± SD of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.