Figure 7.

Protective actions of LGG on junction proteins were mediated by pathways independent of MAPK/ERK signaling cascade in human colonoids. Human colonoids were treated with LGG-CM or EGF (100 ng/ml) for 90 min in the presence or absence of 1-h pretreatment with an EGFR inhibitor, AG1478 (200nM). In separate experiments, human colonoids were treated with IFN-gamma (200 ng/ml) in the absence or presence of LGG. The protective action of LGG was examined in the presence of AG 1478 (200nM). Cellular lysates were prepared and immunoblotted for P-EGFR, total EGFR, P-ERK, total ERK, occludin, ZO-1 and GAPDH. Bands of GAPDH were used as control for an equal protein loading. The optical density is expressed in arbitrary units normalized against a control sample. Data in histograms represent means ± SE; n = 5 in each group (control group vs EGF group *p<0.05, control group vs LGG group #p<0.05, control group vs IFN-gamma group, §p<0.05).