|Our specimens had been fixed with formalin from 24 to 48 h at room temperature and treated with routine processing as in a previous study (12). Ten 4-mm serial sections were prepared for staining and were incubated with primary antibodies for 2 h. Immunohistochemistry was performed using a Discovery Auto-Stainer with automated protocols (Ventana Medical Systems, Inc.; Roche) as previously described (13). The average number of PCNA-positive cells from case 1 to case 23 were counted in 10 random microscopic fields at 400 magnification.
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