“…The 4 µm-thick, FFPE slices were de-paraffinized and rehydrated using a xylene and alcohol solution. Immunostaining was performed using automated instruments (Ventana Benchmark XT (Ventana Medical Systems) and or Dako Omnis (Dako, Carpinteria, CA, USA)) [1,[18][19][20][21][22][23][24][25][26]. After antigen retrieval, the slices were incubated with primary antibodies including pan-cytokeratin (pan-CK; 1:600, clone AE1/AE3, Dako), CK7 (Dako, 1:100, clone OV-TL 12/30, Dako), CK20 (1:100, clone Ks20.8, Dako), caudal type homeobox 2 (CDX2; 1:400, clone EPR2764Y, Cell Marque, Rocklin, CA, USA), paired box 8 (PAX8; 1:50, polyclonal, Cell Marque), estrogen receptor (ER; 1:150, clone 6F11, Novocastra, Leica Biosystems, Newcastle Upon Tyne, UK), progesterone receptor (PR; 1:100, clone 16, Novocastra), p53 (1:300, clone DO-7, Novocastra), p16 (prediluted, clone E6H4, Ventana Medical Systems), p63 (1:50, clone 4A4, Dako), and GATA-binding protein 3 (GATA3; 1:150, clone L50-823, Cell Marque).…”