Fig. 5

BMSCs undergo cytoskeletal rearrangements and re-localize YAP in the cytosol when treated with the combination RBA plus ASC in co-culture with leukemic cells. A The right panels show representative images of MS-5 cells treated in co-culture with MOLM-13 for 72 h, then stained with phalloidin to detect F-actin (orange). Cells were counterstained with Hoechst to identify nuclei (blue). The right panels show the same analysis, but MS-5 cells were treated with the combination RBA plus ASC in the absence of the MOLM-13 cells. B YAP protein levels in MS-5 cells, treated in co-culture with MOLM-13 cells (upper panel) or in mono-culture (lower panel), were evaluated by western blot analysis, upon nucleus-cytosol fractionation. Lamin A/C was used as nuclear marker and stain free protein detection technology (BioRad) for total protein normalization. The graphs report the average ratio of YAP amount in the cytosol over that in the nucleus (paired Student’s T test). C Single confocal images of a Z stack, taken in the center or in the apical portion of MS-5 cells, stained as in (A), to examine the actin cap. D The violin plot reports the measurements of MS-5 nuclear thickness, obtained by the confocal analysis shown in (C) and in Supplemental Fig. 6. E PI exclusion assay showing cell death of MOLM-13 leukemic cells co-cultured for 72 h with MS-5 cells silenced (siYAP) or not (siNC) for YAP protein (two-way ANOVA). F CTGF and GLS mRNA expression levels of MS-5 cells as mono- or co-culture with MOLM-13 cells, upon 72 h of treatment (two-way ANOVA)