Effect of 10 nM epidermal growth factor (EGF) administration to SF media on protein expression of essential autophagic regulators in IEC-6 cells. A: IEC-6 cells were incubated in S media (n = 9), SF media (n = 9), and SF media + 10 nM EGF (SF+EGF, n = 9) for 12 h. Samples were collected and analyzed by Western blotting for Beclin 1. Densitometric values were normalized to β-actin and expressed as a relative level to S values. Western blot images are representative for each group. #P < 0.01 vs. S or SF+EGF. B: representative picture of Western blot for LC3I and LC3II proteins. Densitometric values were normalized to β-actin and expressed as a relative ratio between LC3II and LC3I isoforms. C: transmission electron microscopy (TEM) images of IEC-6 cells. Normal morphology was observed in IEC-6 cells incubated in medium containing S. IEC-6 cells grown in SF medium show the presence of many autophagic compartments that are electron dense, indicating the presence of degraded cytoplasmic material (black arrows). In contrast, TEM images from the cells treated with SF+EGF show normal morphology. M, mitochondria; N, nucleus.