Fig. 6

Effect of NF-YAs and NF-YAl overexpression on tumor growth and cell dissemination in vivo. A Weight (mg) of xenograft tumors at 5 weeks from s.c. inoculation of Empty, NF-YAl and NF-YAs PC3 cells into SCID Hairless Outbred (SHO®) mice. Data represent mean ± SEM (one-way ANOVA with Fisher’s LSD test: *p < 0.05, ns, not significant, n = 8). B Rate of growth (%) of Empty, NF-YAs and NF-YAl xenograft tumors at the indicated time points following s.c. inoculation of transduced PC3 cells. Data represent mean ± SEM (two-way ANOVA with Holm-Sidak’s test: *p < 0.05, n = 8) C Representative H&E-stained FFPE sections of 5 weeks xenograft tumors at low (left panel) and high (middle panel) magnifications. Immunohistochemical detection of Ki67 in xenograft tumors (right panel). D Western blot of total extracts from PC3 tumor xenograft with the indicated antibodies. Tubulin has been used as loading control. Quantification of band intensities was performed with ImageJ software and relative phospho-AKT(Ser473) levels are indicated, after normalization to Tubulin and AKT expression (E) Representation of the incidence of spontaneous cell dissemination to lung tissue in mice after 5 weeks from s.c. injection of Empty, NF-YAl, NF-YAs PC3 cells. Cell dissemination has been identified by the detection of human genomic DNA through Alu-qPCR in mouse lung tissues. F RT-qPCR analysis of the indicated transcripts in xenograft tumors harvested following 5 weeks from s.c. injection. Rps20 and b-Actin were used as reference genes and normalized mRNA levels are reported as fold change vs Empty biological group, arbitrarily set at 1. Data represent mean ± SEM (one-way ANOVA with Tukey’s test: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, n = 5)