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. 2020 Jan 8;10:1503. doi: 10.3389/fphar.2019.01503

Figure 3.

Figure 3

Eth induces autophagy through the AMPK/mTORC1 signaling. (A): MCF-7, MDA-MB-231, or MDA-MB-436 cells were treated with increasing concentrations of Eth for 24 h. Western blot was performed using antibodies indicated. (B): MCF-7, MDA-MB-231, or MDA-MB-436 cells were treated with 2.4 µM, 2.0 µM, or 2.0 µM Eth for the indicated times, and cell lysates were subjected to western blot assay. (C): MCF-7, MDA-MB-231, or MDA-MB-436 cells were treated with Eth (2.4 µM, 2.0 µM, or 2.0 µM) and/or Compound C (CC, 25 µM) for 24 h, and western blot was performed using antibodies indicated. (D): MDA-MB-231 or MDA-MB-436 cells transfected with pQCXIP-GFP-LC3 plasmid were treated with Eth (2.0 µM) and/or CC (25 µM) for 24 h, and assessed by immunofluorescence analyses. Scale bar = 20 µm. (E): Graph shows quantification of LC3-positive punctate cells in (D). (F): MCF-7, MDA-MB-231, or MDA-MB-436 cells were treated with Eth (1.6 µM, 1.0 µM, or 1.6 µM) and/or metformin (Met, 10 mM) for 24 h, and western blot was performed using antibodies indicated. (G): MDA-MB-231 or MDA-MB-436 cells transfected with pQCXIP-GFP-LC3 plasmid were treated with Eth (1.0 µM or 1.6 µM) and/or Met (10 mM) for 24 h, and assessed by immunofluorescence analyses. Scale bar = 20 µm. (H): Graph shows quantification of LC3-positive punctate cells in (G). *P < 0.05, **P < 0.01.