neg C Z Z 15 Multinodular goiter N/C N/C F Z 16 Follicular adenoma C N F Z 17 Multinodular goiter N/C N/C F Z 18 Multinodular goiter N/C N/C F F 19 Papillary cancer & Hashimoto C C F Z C = cytoplasmic; N = nuclear; F = focal; Z = zonal; ND = not determined; neg. = Tozasertib manufacturer negative Biopsy tissues used for immunohistochemical analyses were obtained from normal tissue adjacent to diseased areas. Bucladesine samples were immediately

frozen in liquid Nitrogen and stored at -80°C. On the day of analysis, tissue samples were gradually set to the temperature of -30°C for cryostat procedure. Seven sections were cut from each sample. The immunoperoxidase method was applied with Vector reagents utilizing the following

primary antibodies: a) the anti-p53 polyclonal antibody CM-1 (Novocastra Laboratories Ltd) dilution 1:1000, b) the anti-STAT3 polyclonal antibody C-20 sc-482 clone (Santa Cruz Biotechnology) dilution 1:1000, c) the anti-CK19 monoclonal antibody b170 (Novocastra Laboratories Ltd) dilution Caspase Inhibitor VI ic50 1:100, d) the anti-gp130 polyclonal antibody H-255 (Santa Cruz Biotechnology) dilution 1:250. The staining pattern was evaluated in epithelial cells both in terms of percentage of stained cells and staining intensity. In terms of percentage of stained cells, samples were classified as diffuse, zonal, focal and negative when the % of positive cells was >50%, between 10-50%, <10% and 0%, respectively. In terms of staining intensity, samples were subdivided into three categories: 1 + (low), 2 + (intermediate)

and 3 + (high). Results The results of immunohistochemical analyses are shown in Table 1. Except for case number 8 (multinodular goiter) that was negative for both STAT3 and p53 expression, and case number 14 (papillary SPTBN5 carcinoma) which was negative for STAT3, a diffuse pattern with an intermediate intensity in both nuclear and/or cytoplasmic localizations was observed in all the samples analyzed. An exclusive cytoplasmic localization of STAT3 was seen in 7 cases while a nuclear/cytoplasmic staining was detected in 10 cases. As for p53, three cases displayed an exclusive nuclear staining, 8 cases showed an exclusive cytoplasmic localization, 7 cases showed a nuclear/cytoplasmic positivity [Figure 1] and one case displayed no staining. gp130 staining was negative in two cases (3 and 8) while a zonal or focal membrane and cytoplasmic staining distribution of intermediate intensity (2+) was observed in most of the cases [Figure 2]. Cases 7, 15 and 19 showed an intense (3+) staining. Cytokeratin 19 (CK19) could not be determined in case 3, while 7 samples were negative, 8 showed a focal and 3 a zonal cytoplasmic distribution of intermediate intensity (2+).