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. 2017 Nov 14;8(64):108006–108019. doi: 10.18632/oncotarget.22444

Figure 5. ROS is involved in CD-induced changes in expression of certain NOX/DUOX isoforms and modulation of TSC/Rheb/mTOR signaling axis to regulate autophagy and apoptosis in NPC-BM and NPC-039 cells.

Figure 5

Cells were pretreated with or without 10 mM NAC for 4 h, and then cultured with or without CD (8 μM) for 24 h. Equivalent amounts of total cell lysates were subjected to Western blot analysis. (A) NOX1, NOX2, NOX3, NOX4, NOX5, DUOX1, and DUOX2 were detected in cell lysates by Western blot. (B) The relative density of the bands was quantified by densitometry analysis. The histograms illustrate the relative expression levels after normalization to β-actin. (C) The TSC1, TSC2, Rheb, p-mTOR (Ser2448), mTOR, LC3-I, LC3-II, and p62 were analyzed by Western blot with their respective antibodies. (D) The histograms illustrate the relative expression of the indicated proteins normalized to β-actin. (E) Levels of cleaved caspase-3, cleaved caspase-9, and cleaved PARP were analyzed by Western blot. (F) The relative density of the bands is shown in the histogram. Values represent the mean ± SEM. from three determinations. *P < 0.05, compared with the control (0 μM). #P < 0.05, compared cells treated with CD (8 μM) alone.