Fig. 2
From: Mutant p53s generate pro-invasive niches by influencing exosome podocalyxin levels
Exosomes from mutant p53-expressing cells influence DGKα-dependent integrin trafficking and cell migration in p53 null cells. a H1299-p53−/− ‘recipient’ cells were pre-treated for 72 h with exosomes collected from H1299-p53−/−, H1299-p53R273H or H1299-p53R175H ‘donor’ cells, or were left untreated. Recipient cells were then trypsinised and re-plated. Seventy-two hours following re-plating, recycling of integrin α5β1, cMET and TfnR was determined. Recipient cells were treated with R59022 (10 μM) or DMSO control as indicated. Values are mean ± SEM, n = 6; ***red versus blue, and ***green versus black are p < 0.001 ANOVA. b H1299-p53−/−, H1299-p53R273H or H1299-p53R175H ‘recipient’ cells were pre-treated with exosomes collected from H1299-p53−/−, H1299-p53R273H or H1299-p53R175H ‘donor’ cells, or were left untreated as indicated. Cells were then re-plated and the speed, ΔPersistence and ΔFMI of migration into scratch-wounds determined as for Supplementary Figure 1a–d. Values are mean ± SEM; n > 195 cells from three individual experiments; *** in the right panel, and ***yellow versus green and ***purple versus green in the left panel are p < 0.001, Mann–Whitney. c H1299-p53tetON cells were incubated in the presence or absence of doxycyclin and induction of wild-type p53 was confirmed by western blotting. Exosomes from these cells were incubated with H1299-p53−/− cells, the cells re-plated and the speed, ΔPersistence and ΔFMI of migration into scratch-wounds determined as for b. Values are mean ± SEM; n > 110 cells; *** are p < 0.001, Mann-Whitney. d H1299-p53−/− recipient cells were pre-treated with exosomes derived from H1299-p53R273H donor cells. Recipient cells were then transfected with siRNAs targeting RCP (siRCP) or a non-targeting control (siNT), and the characteristics (∆Persistence, ∆FMI and speed) of their migration into scratch-wounds was determined in the presence and absence of a DGK inhibitor (R59022; 10 μM) or DMSO control. Values are mean ± SEM; n > 273 cells; *** in right panel, and ***green versus black and ***purple versus yellow are p < 0.001, Mann-Whitney test
