IHC analysis of Ep-ICD and EpEx subcellular localization in thyroid tissues.

<p>The IHC score cut-off values for positivity were defined as ≥2 for nuclear Ep-ICD positivity; ≥5 for cytoplasmic Ep-ICD positivity and ≤5 for loss of membranous EpEx expression. ESLI cut-off ≥6 was used to determine ESLI positivity for distinguishing PTC from benign cases; and a cut-off value of >10 was used to determine ESLI positivity for distinguishing aggressive PTCs from non-aggressive PTCs.</p

Clinicopathological characteristics of thyroid carcinoma patients.

<p>Clinicopathological characteristics of thyroid carcinoma patients.</p

Kaplan Meier survival analysis for ESLI.

<p>Kaplan Meier survival analysis showing significant association with reduced disease free survival (DFS) in ESLI positive PTC patients (p = 0.039) with a mean DFS = 133 months compared to ESLI negative patients with a mean DFS = 210 months.</p

Comparison of IHC scores for Ep-ICD, EpEx and ESLI in thyroid tissues.

<p>Mann-Whitney test <i>p</i> value for benign vs. PTC: cytoplasmic Ep-ICD <i>p</i> = 0.000; nuclear Ep-ICD <i>p</i> = 0.002 and ESLI <i>p</i> = 0.000. <i>p</i> value for Aggressive vs. Non-aggressive PTC: cytoplasmic Ep-ICD <i>p</i> = 0.000; nuclear Ep-ICD <i>p</i> = 0.000 and ESLI <i>p</i> = 0.000.</p

Immunohistochemical analysis of EpEx and Ep-ICD expression in papillary thyroid carcinomas and benign tissues.

<p>The representative photomicrographs show immunostaining of EpEx and Ep-ICD in paraffin-embedded thyroid benign nodule goiters, non-aggressive PTC and aggressive PTC tissues. Strong membranous EpEx immunostaining was observed in benign cases (A) and non-aggressive PTC tissues (C); reduced staining of membrane EpEx was observed in aggressive PTC cases (E, G). The benign thyroid nodules and non-aggressive PTC (D) showed predominant cytoplasm localization of Ep-ICD and no detectable nuclear Ep-ICD staining (B, D), while the aggressive PTC cases showed strong nuclear and cytoplasmic Ep-ICD accumulation (F, H). M, membrane staining; C, cytoplasmic staining; N, nuclear staining; Loss of M, loss of membrane expression. Original magnification × 400.</p

Fluorescence immunostaining with anti-EpEx and anti-Ep-ICD antibodies in aggressive and non-aggressive papillary thyroid carcinomas.

<p>Secondary antibodies are FITC-anti-mouse (green) and TRITC-anti-rabbit (red). A-F images from a non-aggressive PTC; G-L Images from an aggressive PTC. A,G) EpEx; B,H) DAPI; C, I) Ep-ICD; D) EpEx and DAPI (A & C merged); E) Ep-ICD and DAPI (B & C merged); F) EpEx, Ep-ICD, and DAPI (A, B, C merged). J) EpEx and DAPI (G & I merged); K) Ep-ICD and DAPI (H & I merged); L) EpEx, Ep-ICD, and DAPI (G, H, I merged). M, Membranous staining; C, Cytoplasm staining; N, Nuclear staining. Original magnification × 400.</p