Fig. 5

Migratory and invasive potential of HT29 cells is reduced upon MIEN1 knockout. (A) Representative brightfield microscopy images of spheroids from days 0 to 4 in HT29 WT and KO cells. The number of spheroids formed each day was quantified manually and is represented by a bar graph. A multiple comparison test was performed to analyze the statistical differences between the groups, with n = 3 in each group. The area of spheroids on each day was measured using ImageJ software and is represented on the bar graph, with n = 3 (top right-end) Scale bars: 100 µm. (B) Representative two-dimensional migration assay images depicting the scratch at the time points mentioned above. The percentage of wound closure was measured using ImageJ software, and statistical analysis between the groups for each day was performed using two-way analysis of variance, with n = 5. (C) Representative image of the Boyden chamber insert used for the transwell invasion assay of HT29 WT and KO cells. The bar graph represents the quantified number of cells in each image captured and averaged. The two-tailed unpaired t-test was used to compare the groups, with n = 3 in each group. GraphPad Prism 10 was used to generate graph and perform statistical analysis with *, p < 0.05; **, p < 0.005; ***, p < 0.0005, ****, p < 0.0001 (microscope software details, magnification or zoom). MIEN1, migration and invasion enhancer 1; KO, knockout; WT, wild-type.