Fig. 4

WDR5 silencing significantly reduced breast cancer metastasis. a) Metastasis free survival in breast cancer patients (n = 295) was calculated in WDR5-low and WDR5-high groups divided at median of gene expression. Statistic comparison was performed using log-rank test. b) Representative IHC staining of WDR5 expression in normal breast tissues (n = 11) and Metastatic Breast Cancer (MBC) PDXs (n = 8) is reported. Scale bar 100 μm. Box plots represent WDR5 expression quantified by using Fiji tools for DAB positivity. Statistical significance was determined using an unpaired Student t test (***: P < 0.001). c) Effect of WDR5 silencing on metastasis formation in vivo in MDA-MB-231. Luciferase-transduced cells were infected to silence WDR5 or a control vector (SCR) and then transplanted in NSG mice (n = 12 per group). Metastases at distant organs were detected by using bioluminescence technique in vivo after resection of primary tumors as reported in Fig. d) Dot plots represent luminescence expressed in radiance of photons detected at metastatic sites in SCR and shWDR5 groups. Values were calculated by IVIS Illumina Software and compared by applying an unpaired Student t test (P = 0.0010). e) Differences among SCR and shWDR5 groups in terms of percentage of survival were calculated by using Log-rank test (n = 7; P = 0.0003). f) RT-PCR was performed in SCR and shWDR5 MDA-MB-231 cells. Bars represent Log2FC of ∆∆CT values of shWDR5 to SCR of WDR5-target genes, classified on the basis of their biological functions