Fig. 5: Everolimus induces autophagy in MDST8. | Cell Death & Disease

Fig. 5: Everolimus induces autophagy in MDST8.

From: Everolimus and plicamycin specifically target chemoresistant colorectal cancer cells of the CMS4 subtype

Fig. 5

A MDST8 and C LoVo cells stably expressing GFP-LC3 were treated with plicamycin (PLI; 25, 50, and 100 nM), everolimus (EVE; 10, 100 nM and 1 µM) or torin1 (TOR; 0.1 μM) for 6 h 24 h and 48 h. After fixation and nuclear staining with Hoechst 33342, the images were acquired by confocal microscopy. Representative images are depicted for each cell line. The scale bar equals 20 μm. B, D GFP- LC3 dots area were quantified. For each assessed parameter and cell line, data were normalized to the untreated control. Data represent means ± SD. Each condition was compared to the untreated control by means of a paired Student’s t test (**P < 0.01, ***P < 0.001). E–G Human colon cancer MDST8 or LoVo cells were treated with EVE (0.1 μM) or PLI (50 nM) for 72 h. TOR (300 nM) was used for 6 h as a prototypical autophagy inducer. SDS–PAGE and immunoblot were performed, band intensities of LC3-I, LC3-II, p62, and β-actin (ACTB) were assessed, and the ratio LC3-II/ LC3-I (F) and p62/ACTB (G) were calculated. Data are means ± SD of three independent experiments (**P < 0.01, ***P < 0.001 versus untreated MDST8 control; ###P < 0.001 versus untreated LoVo control; Tukey’s multiple comparisons test).

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