Fig. 1

Expression of heparanase isoforms in lymphocytes. The lymphocytes were incubated as described for 4 h, at 37 °C. a Expression index (EI) obtained by digital quantification of immunohistochemistry reaction indicating protein level of both heparanases (HPSE and HPSE2). b The values represent mRNA expression of both heparanases, HPSE and HPSE2, using quantitative RT-PCR (qRT-PCR). The Ribosomal Protein L13a (RPL13a) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were reference endogenous genes. The assays were performed with the peripheral blood mononuclear cell fraction obtained from healthy women. T-lymphocytes, lymphocytes obtained from a woman non-affected by cancer. The circulating lymphocytes were incubated for 4 h at 37 °C as indicated, + MCF-7 cells, with MCF-7 cells; + MCF-7 medium, with conditioned medium obtained from MCF-7 cells; + co-culture medium, with conditioned medium from co-culture; + breast cancer plasma, with plasma obtained from breast cancer patient; + healthy woman plasma, with plasma from healthy women. It is important to note that each value represents the median and range of triplicate assays that were performed using lymphocytes obtained from one healthy donor and plasma samples that were collected from three healthy donors, as well as three different patients with breast cancer. RT-PCR reaction was performed in triplicate for each independent sample. *p < 0.0001 and **p < 0.05 by the Kruskal–Wallis test with Dunn auxiliary test. *HPSE and **HPSE2 expression compared to T-lymphocytes without activation assay