IHC staining was conducted with commercially available tissue microarrays (BS17015a and NGL961; Biomax, Rochester, NY, USA) according to previous protocol23 (link), 24 (link). The tissue microarrays were incubated with a polyclonal rabbit anti-human SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm. Sweden) which was diluted in phosphate buffered saline (PBS) at a ratio of 1:20 for 1 h at room temperature, washed 3 times (each for 5 min in PBS), incubated with biotin-labeled secondary immunoglobulin (1:100, DAKO, Glostrup, Denmark) for 1 h at room temperature, washed 3 times, and treated with 3-amino-9-ethylcarbazole substrate chromogen (DAKO) at room temperature to visualize peroxidase activity16 (link). Labeling index was scored accordance with multiplying quantity by intensity. The quantity was defined as Negative: 0, < 25%: 1, 25–75%: 2, and > 75%: 3 The intensity was defined as Negative: 0, Weak: 1, Moderate:2, and Strong: 325 (link).
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