Paraformaldehyde-fixed NSCLC tissue blocks were cut into 5-μm TMA sections and embedded in paraffin. The sections of TMA were then dewaxed in xylene and rehydrated using a graded series of ethanol solutions. Endogenous peroxidase activity was blocked by immersing the sections in a solution of 3% hydrogen peroxide for 30 min. Then, the sections were microwaved in 10 mM citrate buffer (pH 6.0) at 95°C for 20 min to perform antigen retrieval. After being blocked with goat serum for 30 min, the sections were incubated with the primary antibody (anti-Styk1 ab97451,20 (link) anti-AKT1 (phospho S473) ab81283,21 (link) anti-pan-AKT (phospho T308) ab38449,22 (link) anti-GSK3 beta (phospho Y216) ab75745,23 (link) anti-GSK3 beta (phospho S9) ab75814,24 (link) anti-E Cadherin ab4077225 (link),26 (link) from Abcam, Cambridge, UK; and N-cadherin (D4R1H)27 (link) from Cell Signaling Technology, Danvers, MA, USA) at 4°C overnight. After being washed in PBS (phosphate-buffered saline), the sections were incubated with the appropriate horseradish peroxidase (HRP)-labeled goat anti-rabbit/mouse antibodies. Then, the samples were incubated with reagents of the DAB Elite kit (Dako, Denmark), and counterstained with hematoxylin.