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. 2018 Feb 14;9(2):241. doi: 10.1038/s41419-017-0255-5

Fig. 6. Inhibiting autophagy enhanced NBT-induced apoptosis.

Fig. 6

a MCF-7 cells were cotreated with 12 μM NBT and 40 μM CQ for 24 h compared with control or NBT and CQ treatment alone, and apoptosis was examined by using annexin V with PI staining with flow cytometry. b MCF-7 cells were coexposed to NBT (12 μM) and CQ (40 μM) in the presence or absence of 3-MA (5 mM) for 24 h. Total cellular extract of MCF-7 cells was prepared and subjected to Western blot by using antibodies against CASP9 and PARP. c Total cellular extract, mitochondrial fraction, and cytosol fractions of MCF-7 cells was prepared and subjected to Western blot by using antibodies against Bcl-2, Bax, and Cyt C (mitochondrial fraction and cytosol fraction). d Total cellular extract of MCF-7 cells was prepared and subjected to Western blot by using antibodies against DRP1, Fis1, and Mfn2. e Representative TEM images of MCF-7 cells. MCF-7 cells were exposed to NBT (12 μM) in the presence or absence of CQ (40 μM) for 24 h. N nucleus; M mitochondria; red arrows indicate autophagic vacuoles (scale bars: 2 μm, 500 nm). f Total cellular extract of MCF-7 cells cotreated with 40 μM CQ and 12 μM NBT or 0.25 μM Rapa for 24 h compared with control or NBT, Rapa, and CQ treatment alone was prepared and subjected to Western blot by using antibodies against Atg5, LC3B, and p62. Data represent mean ± SD of three independent experiments