Figure 3

HOXA9 induces P-cadherin expression in EOC cells. (A) Western blot analysis of HOXA9 and cadherin levels in control and HOXA9-knockdown SKOV3ip lines and in vector-control and HOXA9-transfected SKOV3-Par lines. The 34 kD band corresponding to HOXA9 is indicated by an arrow. (B) qRT-PCR analysis of relative CDH3 mRNA levels in SKOV3ip and SKOV3-Par lines. Significance of differences in CDH3 mRNA levels was evaluated by Student t-test. (C) EOC cases from the TCGA Project (n = 567) were stratified according to HOXA9 expression in tumors, where HOXA9 mRNA levels were defined as High (≥ upper quartile) and Low (≤ lower quartile). Significance of differences in CDH3 mRNA levels (log2 scale) between upper and lower quartile sub-groups was evaluated by Mann–Whitney U-test. (D) Schematic representation of the human CDH3 promoter showing locations of the two HOXA9-binding sites (site A: 5′-TCATTTAAAAC-3′ and site B: 5′-TAATTTATTTAATAC-3′). Binding of endogenous HOXA9 in control SKOV3ip cells to these sites was detected by chromatin IP. Negative controls included IP using cells expressing HOXA9 shRNA (shA9-B) and IP with IgG. GAPDH was amplified as an irrelevant gene control.