Fig 1: Identified COMT protein–protein interaction partners displayed by Cytoscape. Proteins involved in regulation of cell adhesion and motility, redox homeostasis, cell cycle, ER organization, protein transport and lipid metabolism according to Uniprot are marked in red, green, purple, orange, blue and yellow, respectively. Size of the nodes represent log2 of the average foldchange of COMT to control. Width of the edges represent p-value for the interaction.
Fig 2: Correlation analysis (A) and hierarchical clustering (B) based on the top correlated gene products from omics analyses could distinguish the MCF7 samples with and without COMT expression.
Fig 3: 2D invasion Transwell assay of COMT overexpressing MCF7 cells (MCF7-COMT) compared to parental MCF7 cells. (A) Box plot of the percentual invasiveness of the cells with and without the presence of chemoattractant (fetal bovine serum, FBS) (n = 9 per cell line). **** p < 0.0001, *** p < 0.001. Boxes are extended from the 25th to the 75th percentile, with a line at the median. The whiskers represent minimal and maximal value in the dataset. The data are cumulative for three independent Transwell experiments. See Fig. S1 for the design of Transwell assay and the calibration curves. See Fig. S3 for results of three individual biologically independent experiments. (B) Western blot analysis of COMT and actin expression in MCF7-COMT and MCF7 parental cells in three biologically independent experiments. See Fig. S2 for raw immunoblotting images.
Fig 4: 3D spheroid invasion assay of COMT overexpressing MCF7 cells (MCF7-COMT) compared to control MCF7-GFP cells. (A) Z-stack images of spheroids formed by MCF7-COMT cells and control cell line have been acquired by confocal laser scanning microscopy for 4 days/96 h after day 0. Data for single representative replicate, see Supplementary Fig. S4 for two remaining replicates and Supplementary Data S1 for full-resolution images. (B) Box plot of the percentual change in volume of the spheroids for all replicates (n = 3 per cell line). Boxes are extended from the 25th to the 75th percentile, with a line at the median. The whiskers represent minimal and maximal value in the dataset. *p < 0.05. (C) Western blot analysis of COMT and actin expression in MCF7-COMT and MCF7-GFP cells in three biologically independent experiments. See Fig. S5 for raw immunoblotting image.
Fig 5: In silico analysis of protein–protein interactions between COMT and SPINT2. Analysis was conducted in STRING database using default settings except minimum required interaction score was set to medium confidence (0.400) and maximum number of interactors was set to no more than 50 interactors in 1st shell. Line thickness indicates the strength of data support.
Supplier Page from Abcam for Anti-COMT antibody