Fig. 3.

RIPK4 regulates VEGF-A expression in BC cells and tissues. a Using DNA microarray experiments, the differentially expressed genes (>twofold, P < 0.001) resulting from transfection with T24-shRIPK4 compared with transfection with the control T24-shNC were categorised according to their functions. The number of genes in each category is bracketed. b Seventeen genes, CDH1, CD82, HPSE, MMP7, MET, TP53, SMAD2, CD44, FN1, FAT1, ITGB3, KISS1, MMP11, MMP13, VEGF-A, TGFB1, and MTSS1, showed more than a twofold mRNA differential expression in T24-shRIPK4 cells compared with that in T24-shNC cells related to migration/angiogenesis. c Western blot analysis showed that RIPK4 silenced by shRNA transfection leads to reduced levels of VEGF-A and increased levels of CD82 in T24 and RT4 cells; whereas ectopic overexpression of RIPK4 by pcDNA-RIPK4 transfection had the reverse effects on VEGF-A and CD82 in BIU87 cells. d Representative immunohistochemistry images showing the high expression of RIPK4 and high expression of VEGF-A in a representative BC tissue sample. Original magnification, ×200. The levels of RIPK4 and VEGF-A are correlated positively (P = 0.005). e Upregulation of VEGF-A is significantly associated with poorer survival in patients with BC