Fig. 3: ITF2357 induces a canonical autophagic process. | Oncogenesis

Fig. 3: ITF2357 induces a canonical autophagic process.

From: Histone deacetylase inhibitor ITF2357 leads to apoptosis and enhances doxorubicin cytotoxicity in preclinical models of human sarcoma

Fig. 3

a Representative images of autophagosomal structures by fluorescence microscopy in HT1080 cells stably transfected with EGFP-LC3B vector (HT1080/EGFP-LC3) and treated with 1 and 5 μM ITF2357 for the indicated times. b Quantification of cells positive for punctate autophagosomal structures treated as indicated in a. The results represent the mean ± SEM of three independent experiments. c Western blot analysis of microtubule-associated protein 1 (LC3B-I/II) conversion and of p62/SQSTM1 protein expression in total cell lysates from HT1080 cell line treated with increasing concentration of ITF2357 for 24 h. Western blots representative of two independent experiments with similar results are shown. d Analysis of the indicated mRNA expression evaluated by qRT-PCR in HT1080 exposed to ITF2357 (1 and 5 μM for 24 h). Results are presented as the mean ± SEM of at least two independent experiments. p values were calculated between untreated and treated cells, (*p < 0.05). e Analysis of cell death evaluated by PI staining in HT1080 cells exposed to ITF2357 for 48 h in absence or presence of Chloroquine (CQ), a late stage autophagy inhibitor, *p < 0.05. The results represent the mean ± SD of three independent experiments

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