Anti pcna

For histology, dissected tissues were immediately fixed in 10% (v/v) formalin and embedded in paraffin. Tumor sections (5 μm) were deparaffinized and rehydrated in xylene and graded ethanol series. Antigen retrieval was performed at 125°C for 3 min by a decloaking chamber (Biocare Medical, Pacheco, CA, USA) in target retrieval solution (Dako, Santa Clara, CA, USA). Antigenic proteins were detected using a Zymed Histostain kit (Invitrogen-Thermo Fisher, Waltham, MA, USA) according to the manufacturer's instructions. The following primary antibodies (1 : 100 dilution) were used: anti-cyclin D1 (#2978), anti-vimentin (#5741), anti-β-catenin (#8480), (all from Cell Signaling Technology, Danvers, MA, USA), anti-keratin 5 (#905504, BioLegend, San Diego, CA, USA), and anti-PCNA (#HPA030522, Atlas Antibodies, Bromma, Sweden). Hematoxylin and eosin (H&E) stainings were performed with automated H&E stainer (ST5010 Autostainer XL, Leica Biosystems, Wetzlar, Germany).

For histology, tissues were fixed in 10% (v/v) formalin, embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined by light microscopy. Antigenic proteins were detected using a Zymed HistoMouse SP Kit (ThermoFisher) according to the manufacturer’s instructions. The primary antibodies used were anti-phospho-histone H3 (Merck Millipore) and anti-PCNA (Atlas Antibodies). Apoptotic cells were assessed using TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) assays (Merck Millipore), and the accumulation of collagen in tissues was monitored using Masson’s trichrome staining (Sigma). All the histochemical analysis comparisons were performed in a manual tissue microarray (Unitma, Seongnam, Korea) containing control and treated samples within the same slide.