Fig. 6

Suppressing ACK1 stimulated an autophagy-like response and activated the AMPK/mTOR pathway in lung cancer cells. A A549 cells were infected with adenoviral vectors carrying mRFP-GFP-LC3 and cultured for 24 h, followed by incubation with AIM-100 (20 μM) or Dasatinib (20 μM) for an additional 12 h. mRFP-LC3 and GFP-LC3 puncta, representing autophagosomes, in A549 cells induced by AIM-100 or Dasatinib (at × 400 magnification). The number of LC3 puncta per cell was quantitated. B A549 cells were treated with Dasatinib (20 μM) for 12 h. Cell lysates were analyzed, and immunoblots were probed with indicated antibodies. C, E) A549 cells were treated with different concentrations of AIM-100 for indicated hours. Influences of AIM-100 on common biomarkers of autophagy C and the AMPK/mTOR signaling pathway E were examined. D, F NCI-H1299 cells were used to investigate the effects of ACK1 overexpression on the biomarkers of autophagy (D) and the AMPK/mTOR signaling (F). G A549 cells were incubated with AIM-100 (20 μM) and/or AMPK inhibitor (Compound C, 20 μM) for 12 h, succeeded by Western blot. *P < 0.05, **P < 0.01, ***P < 0.001