ATF5 inhibits autophagy in BCR-ABL–transformed cells. (A) 32D/BCR-ABL or K562 cells treated with either a NS or ATF5 shRNA were monitored for expression of ATF5 and LC3B by immunoblot analysis. β-actin (ACTB) was monitored as a loading control. The fold change in LC3B-II levels on ATF5 knockdown was quantified by measuring the intensity of the LC3B-II signal in the immunoblots and normalizing to ACTB levels. (B) 32D/BCR-ABL or K562 cells ectopically expressing a GFP-LC3B fusion protein were treated with either a NS or ATF5 shRNA and analyzed by fluorescence microscopy. Representative images are shown. (C) Immunoblot analysis of p62 levels in K562 cells expressing a NS or ATF5 shRNA. (D) K562 cells expressing a NS or ATF5 shRNA were stained with acridine orange in the absence or presence of bafilomycin A1 (Baf A1). (E) Immunoblot analysis of ATF5 and LC3B levels in K562 cells stably expressing FLAG or FLAG-ATF5, and treated in the absence or presence of imatinib (IM; 10μM for 48 hours).