Depleted FLJ10540 attenuates Aurora-A elicited malignant phenotype changes in vitro and in vivo. (A and B) The mRNA and protein expression levels of FLJ10540 were determined by Q-RT-PCR and Western blotting in vehicle or overexpressing Aurora-A transfectants. The cell lysates of FaDu/vehicle-negative control, FaDu/vehicle-siFLJ10540, FaDu/Aurora-A-negative control, or FaDu/Aurora-A-siFLJ10540 transfectants were subjected to immunoblot analysis with anti-FLJ10540, and β-actin antibodies. Data are representative of three independent experiments done in triplicates. (C and D) Using the same panels, the proliferation, migration, and invasion assays were also performed by MTT assay and Transwell chambers. (E) Centrosome numbers were determined in Aurora-A stable cells transfected with negative control or siFLJ10540 in FaDu cells by using gamma-tubulin antibody. (F) Antitumor activity of MLN8237 plus siFLJ10540 in a SAS xenograft model. Nude mice bearing subcutaneously established SAS alone (n = 6), SAS/negative control (n = 6) or SAS/siFLJ10540 (n = 6) xenograft tumors and received the indicated treatments. Tumor growth was monitored and was shown as mean volumes ± SD. The paraffin-embedded tumor tissues were subjected to immunostaining for FLJ10540. Statistical analysis: *p < 0.05, **p < 0.01, ***p < 0.001.