Fig. 6: YB-1 is necessary for the cytoprotective effects of ZO-1 downregulation.

a Cytoprotection induced by the absence of ZO-1 is not maintained in the absence of YB-1. Cell viability, measured by Trypan blue exclusion, of HUVECs transfected with siRNA against CT, ZO-1, YB-1 or ZO-1 and YB-1 and treated or not with arsenite (25 µM; 12 h) (n = 4 independent experiments). b EC migration induced by the absence of ZO-1 is not maintained in the absence of YB-1. Quantification of the percentage of wound closure at 16 h of migration, normalized to the control, of BAECs transfected with siCT, siZO-1, siYB-1 or ZO-1 and YB-1 treated or not with arsenite (10 µM) (n = 9 fields of view per condition). c Increased transcriptomic targets of YB-1 in the absence of ZO-1. Immunoblot analysis of BCL-XL or G3BP1 levels in siCT, siZO-1 and siYB-1 transfected HUVECs with siRNA. Histogram referring to the quantification of BCL-XL or G3BP1 levels relative to β-actin (siCT n = 10, siZO-1 n = 6, siYB-1 n = 5 independent experiments). d ZO-1 influences translation initiation during EC stress independently of YB-1. Immunoblot analysis of serine 51 phosphorylation of eIF2α levels in siCT, siZO-1, siYB-1 or siZO-1 and siYB-1 transfected HUVECs and treated or not with arsenite (500 µM; 30 min). Histogram referring to the quantification of serine 51 phosphorylation of eIF2α relative to total eIF2α (siCT, siCT + arsenite, siZO-1, siZO-1 + arsenite, siYB-1, siYB-1 + arsenite n = 6; siZO-1 + siYB-1, siZO-1 + siYB-1 + arsenite n = 4 independent experiments). a–d Groups were compared using one-way ANOVA followed by Bonferroni’s multiple comparison tests; ns not significant. Data are presented as mean values ± SEM. Source data are provided in the Source Data file.