Fig. 5: Inhibition of autophagy enhances GANT61-induced apoptosis. | Cell Death & Disease

Fig. 5: Inhibition of autophagy enhances GANT61-induced apoptosis.

From: Inhibition of the sonic hedgehog pathway activates TGF-β-activated kinase (TAK1) to induce autophagy and suppress apoptosis in thyroid tumor cells

Fig. 5

A SW1736 cells were treated with the indicated concentrations of GANT61 for 48 h or with 10 μM GANT61 for the indicated lengths of time. B, D SW1736 cells were incubated in the absence or presence of GANT61 (10 μM) minus or plus 5Z (1 or 5 μM) (B) or CQ (5 or 10 μM) (D) for 24 h. Cell lysates were prepared and analyzed for PARP, caspase-8 (C8), cleaved caspase-8 (CC8), caspase-3 (C3), and cleaved caspase-3 (CC3) by western blot. C-PARP cleaved PARP. C, E SW1736 cells were incubated in the absence or presence of GANT61 (10 μM) minus or plus 5Z (1 or 5 μM) (C) or CQ (5 or 10 μM) (E) for 24 h. Single-cell suspensions were stained for propidium iodide (PI) and annexin V followed by flow cytometry. Data in bar graphs are the mean ± SD of three independent experiments. F, G SW1736 cells were transfected with scrambled or TAK1 siRNA (F) or Beclin-1 siRNA (2.5 nmole each) (G). After incubation for 24 h, the cells were left untreated or treated with GANT61 for 24 h. Cell lysates were analyzed for p62, LC3, Beclin-1, TAK1, PARP, C8, CC8, C3, CC3, and β-Actin by western blot. The expression levels were analyzed by quantification of the density of the protein bands with NIH Image-J software and presented as bar graphs. *p < 0.05; **p < 0.01, compared to the untreated control; #p < 0.05; ##p < 0.01, compared to GANT61 (A–E).

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