Aurora-A upregulates FLJ10540 in HNC. (A and B) Flag-tagged FLJ10540 stable clone of FaDu cells and siFLJ10540 transfectants of SAS cells were established. The cell lysates were subjected to immunoblot analysis with anti-Flag and FLJ10540 antibodies. β-actin is as an internal control. Aurora-A mRNA and protein expression levels in FLJ10540-overexpressing and vehicle cells were recognized by Western blotting and Q-RT-PCR. (C and D) Flag-tagged Aurora-A stable clone of FaDu cells and siAurora-A transfectants of SAS cells were established. The endogenous mRNA and protein of FLJ10540 were detected by Western blotting and Q-RT-PCR. (E and F) The luciferase assays were performed to detect promoter activities of FLJ10540 in cotransfected with in a dose-dependent manner of Flag, Flag-Aurora-A, negative control or siAurora-A. The FLJ10540 luciferase activity was normalized to Renilla activity. Data are representative of three independent experiments done in triplicates. Images were quantified with ImageJ software. Statistical analysis: *p < 0.05, ***p < 0.001.