Fig. 2
From: c-Jun N-terminal kinase activation by nitrobenzoxadiazoles leads to late-stage autophagy inhibition
MC3181 and NBDHEX induce accumulation of both autophagic vesicles and the selective autophagy substrate p62. a Morphology of U-2OS cells treated with vehicle (CTRL), 5 μM MC3181 or 5 µM NBDHEX for 48 h, viewed under an Olympus IX50 inverted microscope equipped with a digital camera. Original magnification ×200. b Puncta formation assay on U-2OS GFP-LC3 cells treated with vehicle (CTRL), 5 μM NBDs or 10 μM CQ for 48 h. Cells were photographed using a fluorescence microscope equipped with a digital camera. Original magnification ×400. c Immunoblot analysis for LC3-II and p62 in lysates from U-2OS, MM-B1, MCF-7, COLO 205 and HT-29 cells treated with vehicle (CTRL), 5 μM MC3181 or 5 µM NBDHEX for 48 h. β-Actin was used to ensure equal loading and transfer of samples. LC3-II/β-actin and p62/β-actin densitometric ratios are also reported: those of MC3181- and NBDHEX-treated samples were expressed in proportion to those of CTRL samples, which were arbitrarily set, equal to 1