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. 2017 Jun 8;8:351. doi: 10.3389/fphar.2017.00351

Figure 2.

Figure 2

Tetrandrine (Tet) induces autophagy in multiple cell lines. (A) A panel of cancer cells lines expressing GFP-LC3 were treated with the indicated compounds at 5 μM for 16 h. (B) Both ATG7+/+ wild-type and ATG7−/− deficient (KO) MEFs were transiently transfected with the GFP-LC3 plasmid for 24 h and then treated with DMSO (Ctrl) or 5 μM Tet for 16 h. The cells were then fixed for fluorescence imaging and cells counting. Magnification: x63. (C) Representative electron micrographs showing the ultrastructures of MCF-7 cells treated with Tet (5 μM) for 0–48 h. Magnification: x5600. Arrows, double-membraned autophagosomes. Magnification: x24000. (D) MCF-7 cells were treated with Tet (5 μM) and lysosomal protease inhibitors (E64d and pepstatin A) (10 μg/mL each), either alone or in combination, for 0–24 h. Cell lysates were analyzed by western blot for LC3 conversion. (E) Induction of p62 by Tet (5 μM) in MCF-7 cells. Cell lysates were analyzed by western blot for p62 and β-actin respectively. The results are representative of three independent experiments.