Journal: Oncogene

Article Title: SUMOylation and SENP3 regulate STAT3 activation in head and neck cancer

doi: 10.1038/onc.2016.124

Figure Lengend Snippet: NNK induces STAT3 phosphorylation and increase of SENP3 protein levels in HNC cells in an ROS-dependent manner. ( a ) The levels of STAT3 phosphorylation at tyrosine 705 (in brief, pY705-STAT3) were determined by IB in Hep-2 cells exposed to 1 μ m NNK and 20 ng/ml IL-6 respectively for the indicated time (left), and in KB or HN30 cells exposed to 1 μ m NNK for indicated time (right). ( b ) The pY705-STAT3 levels were determined by IB in Hep-2 cells exposed to 1 μ m NNK in the presence or absence of 1 μg/ml neutralized antibody against IL-6 (left) and 10 μ m JAK inhibitor (right) for the indicated time. Anti-IL-6 antibody and JAK inhibitor were pre-administered for 1 h. ( c ) The pY705-STAT3 levels were determined by IB in Hep-2 cells exposed to lower doses of NNK (0.01 μ m ) or IL-6 (2 ng/ml) alone or in combination for the indicated time. ( d ) ROS level was determined by DCFH-DA staining and flow-cytometric analysis in Hep2 cells exposed to 1 μ m NNK for the indicated time. Error bars represent±s.d. for triplicate experiments. *** P <0.001. ( e ) The SENP3 and pY705-STAT3 levels were determined by IB in Hep-2, KB and HN30 cells exposed to 1 μ m NNK in the presence or absence of 5 m m NAC for the indicated time.

Article Snippet: When needed, the anti-oxidant NAC (Sigma, St Louis, MO, USA) was pre-administered for 4 h, and the JAK inhibitor (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-IL-6 (InvivoGen, San Diego, CA, USA) were pre-administered for 1 h before other treatments.

Techniques: Phospho-proteomics, Staining