Fig. 4: UBCS039 promotes autophagic flux in human cancer cells.

a, b Western blot analysis of LC3B-II protein level in H1299 (a) and HeLa (b) cell lines treated with 75 µM UBCS039 in combination or not with 3-MA (1 mM) or CQ (25 µM) for 24 h. LC3B-II levels were quantified by densitometric analyses using ImageJ software and the relative levels of LC3B-II were expressed in the histograms as fold changes of treated versus untreated samples, after HSP70 normalization. Results represent mean ± SD of three independent experiments. *P < 0.05; ***P < 0.001. c H1299-ptf-LC3 cells were treated with 75 µM UBCS039 for 72 h or with 25 µM CQ for 24 h alone or in combination. Representative images of fluorescence microscopy analysis of yellow- and red-fluorescence EGFP-LC3B punctate vesicular structures, indicative of autophagosomes and autolysosomes accumulation, respectively. d Representative images of co-immunofluorescence analysis of lysosomal marker LAMP-2 and EGFP-LC3B punctate vesicular structures in H1299 cells expressing EGFP-LC3 treated or not with 75 µM UBCS039 for 72 h. c, d Olympus AX70 microscope, ×100 magnification. Scale bar indicates 10 µm