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. 2020 Jul 7;8:558. doi: 10.3389/fcell.2020.00558

FIGURE 5.

FIGURE 5

TGFβ does not modify the effects of EVs either in EGF-dependent or in EGF-independent 3D models. (A) The percentage of active caspase-3+ cells in CRC patient-derived organoid lines in the presence or absence of AREG, control, or TGFβ-treated NCF-derived EVs. (B) The effect of NCF-derived EVs on CRC organoids. NCFs were pre-treated with TGFβ (TGFβ EV) or they were cultured untreated (Ctr EV) before collecting EVs. (C) Detecting AREG on EVs isolated from control (Ctr EV) or TGFβ-treated NCFs (TGFβ EV) and bound to latex beads (flow cytometry). (D,E) The percentage of KI67+ cells in SW1222 cell-derived colonies in the presence of the indicated treatments. Representative confocal microscopic images (D) and their quantification (E). EVs were isolated from control or TGFβ-treated (4 days) NCF cultures. Kruskal–Wallis and Dunn’s multiple comparison tests were used (A,B,E). Confocal microscopic images were quantified from three experiments for (A,B,E). Scale bars: 50 μm (D). *p < 0.05, **p < 0.01, ***p < 0.005.