Fig. 7

Colocalization analysis of ceramide and DR5 after GT3 treatment in MIA PaCa-2 cells. MIA PaCa-2 cells were treated with dissolution vehicle (a top panels ) or 40μM GT3 (a bottom panels), fixed, and processed for immunofluorescence. Single confocal sections shown in the x-y plane, and 3D reconstructions of the confocal stack in the x–z plane perpendicular to the monolayer, apical side up, show localization of DR5 (green channel), ceramide (red channel), and nuclei using DAPI (blue channel). Intensity correlation analysis of 14 fields, each with an average of 12.5 cells from at least three independent experiments, using the Leica Software were run for control cells (b) and (c) and treated cells (d) and were plotted for both sets of cell populations. The values shown are the averages ± standard deviations obtained from at least three independent experiments. Images are representative of fields analyzed. Graphs illustrating the quantification of relative fluorescence for control cells (c) and treated cells (e) show the merge for ceramide and DR5 channels. Images shown are representative of the fields observed