Anti aurora a

The E6 DNA fragment of HPV16 was obtained by polymerase chain reaction (PCR) from SiHa genomic DNA and ligated to a PCDNA3.1 vector (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) as previously described (23 (link)). The p3XFLAG-E6 expression vector was generated by PCR cloning of the HPV16 PCDNA3-E6 cDNAs, followed by HindIII and XbaI double digestion and insertion into the HindIII and XbaI sites of the pA3F vector (Sigma-Aldrich, St. Louis, MO, USA). Human Aurora A cDNA (a gift from Dr Bingyi Xiao, University of Pennsylvania, Philadelphia, PA, USA) was subcloned into the pcDNA3.1HA (Invitrogen; Thermo Fisher Scientific, Inc.) and pGEX-4T-2 vectors (GE Healthcare Life Science, Chalfont, UK) to produce hemagglutinin (HA)-tagged and glutathione S-transferase (GST)-tagged plasmids. The antibodies used in the present study were the mouse monoclonal anti-FLAG M2 (catalog no. F1804; Sigma-Aldrich), anti-HA (catalog no. H3663; Sigma-Aldrich), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-Aurora A (catalog no. 610939; BD Biosciences, San Jose, CA, USA) and anti-HPV16 E6 C1P5 (catalog no. sc460; BD Biosciences) antibodies (at 1:1,000 dilution for western blotting).