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Fig. 3 | BMC Cancer

Fig. 3

From: Anti-tumor effects of everolimus and metformin are complementary and glucose-dependent in breast cancer cells

Fig. 3

Metformin and everolimus do not influence autophagy and ROS production. a MCF-7 and MDA-MB-231 cells stably expressing LC3-GFP cultured in 11 mM and 2.75 mM glucose-containing medium were treated with indicated concentrations of 10 nM everolimus and/ or 5 mM metformin for 48 h. 20 μM bafilomycin was added 1 h prior to evaluation of the assay. Fluorescent GFP-LC-3 foci per individual cell were counted. Data are presented as mean ± SD of three different experiments. b MCF-7 and MDA-MB-231 cells cultured in 11 mM and 2.75 mM glucose-containing medium were treated with everolimus (10 nM) or metformin (5 mM) for 48 h. Western Blotting for LC-3 revealed that there is no increased LC-3 cleavage after either treatment. c MCF-7 cells were treated with 250 μM H2O2 for 1 h. Hydrogenperoxide induces ROS production in MCF-7 cells, as demonstrated by increase in cellular H2DCF fluorescence. d MCF-7 and MDA-MB-231 cultured in 11 mM or 2.75 mM glucose-containing medium were treated with indicated concentrations of everolimus or metformin for 48 h, and stained with 10 μM H2DCF. Everolimus and metformin do not induce significant changes in ROS levels compared to untreated cells. Data are presented as mean ± SD of three different experiments

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