Fig. 4

FWGE-induced autophagic activity in the colon carcinoma cell line HRT-18. FWGE-treated HRT-18 cells showing intracellular vacuoles 24 h after start of incubation. The vacuoles increased in size with increasing incubation time (a). Presence of endogenous LC3-II in HRT-18 cells with and without FWGE treatment at different incubation times (b). LC3-I (approximately 17 kDa) is the cytosolic form of LC3, which is converted into the active, membrane-bound form LC3-II (approximately 14–15 kDa) during the autophagy process. β-actin was used as a loading control (42 kDa). The shift from LC3-I to LC3-II is evident following FWGE treatment. Western blot results are shown for one trial representative of three independent experiments (c). The LC3-II/LC3-I ratio was calculated based on densitometry analysis (ImageJ 1.3.4 s downloaded from the National Institutes of Health (NIH), Bethesda, MD, USA) of LC3-I and LC3-II bands for three independent experiments (***P < 0.001, *P < 0.05 in comparison to FWEG-untreated cells)