Figure 1
From: MAGI1 inhibits the AMOTL2/p38 stress pathway and prevents luminal breast tumorigenesis
MAGI1 impairment induces tumorigenic phenotypes in epithelial cells. (A) MTT assay (OD 560 nm) representing 2D cell growth of MCF7shMAGI1 as compared to MCF7shLuc cells. Bars represent mean ± Standard Deviation (SD; n = 10 wells as replicates) of a representative experiment (out of 3). Unpaired two-tailed Student's t-test; **p < 0.01; ***p < 0.001. (B) Cell cycle phases of MCF7shLuc and MCF7shMAGI1 cells assessed after BrdU incorporation and analyzed by flow cytometry, showing an increased proportion of cells in S phase at the expense of G0/G1 after MAGI1 invalidation. The pie chart also showed no major difference in the percentage of apoptotic cells in SubG1 phase (around 2% in shLuc and 1% in shMAGI1 cells). (C) Left: quantification of colony numbers of MCF7shMAGI1 cells grown in anchorage independent conditions (soft agar assay) and represented as fold increase compared to MCF7shLuc cells. Data are presented as the means ± SD (n = 3). Unpaired two-tailed Student's t-test; ***p < 0.001. Right: western blot analysis of whole protein extracts issued from the cells used in the soft agar assays and showing the relative amounts of MAGI1 in the different cell lines; Tubulin was used as a loading control. Uncropped blots can be found in the Supplementary Information. (D) Top: quantification of mammospheres represented as total area issued from MCF7shMAGI1 cells and shown as fold increase compared to MCF7shLuc cells, starting from 200 cells. Data are presented as the means ± SD (n = 4). Unpaired two-tailed Student's t-test; ***p < 0.001. Bottom: images of mammosphere formation assay showing a representative experiment.
