Formalin-fixed paraffin-embedded tissues of 30 surgical specimens from patients with prostate cancer, treated with radical prostatectomy were retrieved from the archives of the Department of Pathology, University Hospital of Alexandroupolis. Three μm thick tissue sections were deparaffinized, and antigen retrieval followed by heating in a dry oven for 30 min at 80 °C, in the EnVision FLEX Target Retrieval Solution pH 9.0 (DAKO, Glostrup, Denmark). Endogenous peroxidase was quenched with EnVision Flex Peroxidase Block (DAKO) for 10 min. Next, samples were incubated with the primary rabbit polyclonal anti-PLIN3 antibody (1:100, ab47638, Abcam,UK) for 1 h. Thereafter, sections were incubated with the respective secondary antibody (EnVision Flex/HRP, DAKO) for 30 min at RT. Color was developed after 5 min of incubation time with DAB solution, followed by light counterstaining with Hematoxylin QS (Cat. #H-3404, Vector Laboratories Inc., USA).
Assessment of PLIN3 expression was performed at × 200 magnification. The percentage of cells with absent, weak or strong cytoplasmic expression was recorded per each optical field and the mean value from all optical fields was used to score each case.
Assessment of PLIN3 expression was performed at × 200 magnification. The percentage of cells with absent, weak or strong cytoplasmic expression was recorded per each optical field and the mean value from all optical fields was used to score each case.