Figure 1
Selective BRAF inhibitors induce autophagy in BRAFV600E CRC cells.
(A) HT29 and RKO cells were treated with DMSO and 0.1, 1, or 10 μM PLX4032 for 24 h. LC3-II/LC3-I ratios were determined using Image lab software version 5.0 (Bio-Rad Laboratories). (B) HT29 and RKO cells were treated with DMSO and 0.1, 1, or 10 μM PLX4032 for 24 h. Cells were visualized using a confocal laser microscope (Olympus FluoView FV1000; objective, ×200). Green dots correspond with accumulated LC3 in autophagosomes and nuclei are stained with DAPI (blue). Scale bars indicate 10μm. (C) HT29 and RKO cells were treated with DMSO for 24 h and 10 μM PLX4032 for 2, 6, 12 and 24 h and LC3-II expression and LC3-II/LC3-I ratios were determined using Western blot analyses. (D) HT29 and RKO cells were treated with DMSO for 24 h and 10 μM PLX4032 for 2, 6, 12 and 24 h. Cells were visualized using a confocal laser microscope (Olympus FluoView FV1000; objective,×200). Green dots correspond with accumulated LC3 in autophagosomes and nuclei are stained with DAPI (blue). Scale bars indicate 10μm. (E) HT29 cells were treated with or without 10 μM PLX4032 in the presence or absence of 25 μM CQ for 24 h and Western blot analyses of LC3 turnover were performed.
