Figure 5
Video microscopy images of untransformed (a) and v-Src-transformed MDCK cells (b) cultivated at +35°C in Matrigel for 1, 2, 3, 7 and 8 days, labelled every day with a mixture of a DNA-marker Hoechst (blue), Mitotracker Green (green) and Mitotracker Orange CM-H2TM Ros (orange) for 15 min in the cell culture stove and illuminated under an Olympus Cell M video microscope using a × 40 water immersion objective and appropriate filters for blue, green and red channel. In untransformed MDCK cells lumen formation is visible already on the third day of culture, whereas in v-Src-transformed cells there is no organized structure in the cell cluster, which only increases in size. Apoptotic bodies are visible in Hoechst-stained untransformed MDCK cells in images taken on the seventh and eighth culture days. The average value for the ratio of the intensity of selected regions of interest (ROI) in red and green channel (O/G) is shown below the images and the number of measured ROIs is shown in parenthesis. The ROIs were chosen both from the surface and the interior of the cell cluster and included several cells, but always so that one could distinguish the mitochondria. The statistical t-test showed that in each day the average ratio values in MDCK cells differed statistically significantly from the ones in v-Src-transformed MDCK cells with the confidential limit P<0.05. Bar: 20 μm.
