FIGURE 4.

GYZ stimulates autophagy in NSCLC cells. (A) Immunoblot analysis of LC3 turnover expression in NSCLC cells treated with vehicle or GYZ for 24 h. (B,C) Immunofluorescent analysis of the accumulation of endogenous LC3 puncta in the absence or presence of GYZ (A549 150 μM, H1975 100 μM) for 24 h. Scale bars, 10 μm. (D) NSCLC cells were treated with vehicle, GYZ (A549 150 μM, H1975 100 μM), 3-MA (10 mM), or in combination for 24 h. Immunoblot analysis was used to examine LC3 turnover. (E) NSCLC cells were treated with GYZ (A549 150 μM, H1975 100 μM) with or without CQ (5 μM) for 24 h. LC3 turnover was detected by immunoblotting. (F,G) The colocalization of endogenous LC3 with LAMP2 was quantitated by immunofluorescent analysis in the treatment of GYZ (A549 150 μM, H1975 100 μM) with or without CQ (5 μM) for 24 h. Scale bars, 10 μm.