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. 2010 Jun 10;299(3):G614–G622. doi: 10.1152/ajpgi.00076.2010

Fig. 3.

Fig. 3.

Effect of EGF on autophagosome formation in IEC-6 cells. A, left: GFP-LC3 transfected IEC-6 incubated in S media, SF media, and SF+EGF conditions for 12 h. LC3II was visualized by fluorescent microscopy and observed as green fluorescent puncta. Cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue). An accumulation of autophagosome incorporated LC3II fluorescent puncta (green) was observed in the SF group. Right: enumeration of LC3II puncta per cell is shown (n = 100). #P < 0.01 vs. S or SF+EGF. B: GFP-LC3 transfected IEC-6 incubated for 12 h in one of the following media: S media (complete growth medium containing 10% FBS), SF media, SF+EGF, SF+EGF + 1 μM AG1478 (SF+EGF+AG1478), SF+AG1478, SF media containing 8 nM insulin-like growth factor-I (SF+IGF-I), and SF media containing 10 nM PDGF (SF+PDGF). C: in separate studies, IEC-6 cells were incubated for 4 h in one of the following media: S media (complete growth medium containing 10% FBS), Hanks′ balanced salt solution (HBSS), HBSS containing 10 nM EGF (HBSS+EGF), HBSS containing 10 nM EGF + 1 μM AG1478 (HBSS+EGF+AG1478), HBSS containing 1 μM AG1478 (HBSS+AG1478), HBSS containing 8 nM IGF-I (HBSS+IGF-I), and HBSS containing 10 nM platelet-derived growth factor (HBSS+PDGF). LC3II was visualized by fluorescent microscopy and observed as green fluorescent dots. Cell nuclei were stained with DAPI (blue). The percentage of cells containing LC3II dots was evaluated in 300 cells per sample. *P < 0.01 vs. S or SF+EGF.