Fig. 3: UBCS060, a chemical inactive analog of UBCS039, fails to deaceylate H3K9 and H3K56 and does not induce autophagosome accumulation.

a Western blot analysis of LC3B-II protein level in H1299 cells treated with 75 µM UBCS060 for the indicated times. 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. S.E. short exposure, L.E. long exposure. b Representative images and relative quantification of fluorescence microscopic analysis of LC3B puncta positive cells in H1299 EGFP-LC3B treated as indicated in a. c Representative images and relative quantification of fluorescence microscopic analysis of LC3B puncta positive cells in H1299 EGFP-LC3B transiently silenced for SIRT6 and treated with UBCS039 75 µM for 72 h. Western blot analysis of SIRT6 expression levels. H3 was used as an internal loading control. d Representative images and relative quantification of fluorescence microscopic analysis of LC3B puncta positive cells in H1299 EGFP-LC3B transiently transfected with the wild-type (WT) or with the catalytically inactive form of SIRT6 protein (H133Y) for 48 h. Western blot analysis of SIRT6 expression levels. H3 was used as an internal loading control. b–d Olympus AX70 microscope, ×100 magnification. Scale bar indicates 10 µm. The results represent mean ± SD of three independent experiments. ***P < 0.001