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Figure 5. Fluorescence examination of the resected spheroid tumors. (A) Images of immuno-fluorescent EGFR. H1975 spheroid tumor sections from mice fed with 2 and 4 g/kg of BJ and water control were incubated with EGFR antibody (green) followed by FITC-conjugated secondary antibody treatment before being counter-stained with DAPI (blue) (scale bar, 50 μm). (B) Images of immuno-fluorescent pEGFR^Y1068 H1975 spheroid tumor sections from mice fed with 2 and 4 g/kg of BJ and water control were incubated with pEGFR^Y1068 antibody (green) followed by FITC-conjugated secondary antibody treatment before being counter-stained with DAPI (blue) (scale bar, 50 μm). (C) Images of immuno-fluorescent ALDH1A1 and Nanog H1975 spheroid tumor sections from mice fed with 2 and 4 of g/kg of BJ and water control were incubated with ALDH1A1 antibody (green) followed by FITC-conjugated secondary antibody. The Nanog antibody (red) incubation was followed by TRITC-conjugated secondary antibody before being counter-stained with DAPI (blue) and merged (scale bar, 50 μm). (D) Release of mitochondrial cytochrome c in spheroid tumors. The dissected specimens of H1975 spheroid tumors as treated with BJ (2 and 4 g/kg) and the water control were fixed and incubated with antibody against cytochrome c followed by staining with secondary antibody conjugated with TRITC (red). The slides were counter-stained with Mitotracker (green) and DAPI (blue) before being analyzed by confocal microscopy. The merged images from confocal microscopy with red cytochrome c and green mitochondria signify the appearance of coalescence (yellow), while blue indicates the nucleus (scale bar, 50 μm).