Fig. 1: Loss of Mdm2 or functional impairment of the Mdm2/MdmX complex decreases migration, impairs cell spreading and cell attachment to ECM in a p53-independent manner.

HT1080 p53KO cells were transfected with siRNAs against Mdm2 and siCtrl or treated with the Mdm2 inhibitor, MEL23. A, B Protein levels of Mdm2 and MdmX after (A) transfection using siRNAs (n = 6 samples) as indicated or (B) treatment with MEL23 (7 µM) for 24 h (n = 4 samples). α-tubulin was used as a loading control. C, D Cell migration assay. Representative images (top) and quantification (bottom) of wound scratch migration assay with cells (C) silenced for Mdm2 or (D) treated with MEL23 for 24 h, n = 3 groups. E, F Representative images of the morphology of cells attached to collagen-coated coverslips and quantification of cell area after (E) silencing of Mdm2 or (F) treatment with MEL23 for 24 h, n = 3 groups. Cells stained for actin (orange) and DNA via DAPI (blue). The cell area was quantified and plotted. Graphs show all three independent experimental replicates combined, for each independent experimental replicate a minimum of 100 cells were quantified summing a total of at least 300 cells/condition. G, H Quantification and representative micrographs showing attachment to different ECM components of cells (G) silenced for Mdm2, n = 3 samples or (H) treated with MEL23 for 24 h, n = 4 samples. The graphs shown represent the mean ± SD of independent experimental replicates. More details about the statistical tests used can be found in the Source Data file.