Fig. 5

CD271^high cells display a greater susceptibility to Aβ^(25–35)-induced death. A, CD271 silencing was confirmed by Western blot. B, Melanoma cells were treated with Aβ^(25–35) 40 μmol/L and the amount of dead cells (% sub-G₁) was measured by PI staining at the FACS. C, AnnexinV/PI assay was performed to evaluate the percentage of early and late apoptotic cells. D, CMVTO_EV- and CMVTO_CD271-transfected cells were treated with doxycycline, and proteins were collected to evaluate CD271 induction by Western blot. E, Melanoma cells were treated with doxycycline for 48 hours, followed by Aβ^(25–35) administration. PI staining was performed 24 hours later and the percentage of dead cells was measured by FACS. F, M121224 CD271+ and − cells were seeded as spheroids and treated with Aβ^(25–35). Twenty-four and 144 hours later, brightfield pictures were taken and PI staining (G) was performed. Data represent the mean ± SD of triplicate determinations. H, M130425 wt and CD271 KO cells were injected into the yolk of zebrafish larvae and treated with Aβ^(25–35) ± MEKi. Pictures of wt are the same of Fig. 4C. I, The severity of metastasis was evaluated 4 days later by two blind investigators. J, Probability of survival in zebrafish injected with wt versus CD271 KO cells. Two-way ANOVA was used for statistical analysis. **, P < 0.01; ***, P < 0.001; ****, P < 0.00001; ns, nonsignificant.