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. Author manuscript; available in PMC: 2009 Dec 1.
Published in final edited form as: Int J Cancer. 2008 Dec 1;123(11):2566–2573. doi: 10.1002/ijc.23898

Figure 2.

Figure 2

Heparanase induces VEGF C expression. A. Tumor-derived cell lines were stably transfected with control empty vector (Vo, ◆) or heparanase (Hepa, ■) plasmid and total cell lysates were subjected to heparanase activity assay (upper panel) and immunoblotting applying anti-heparanase (second panel) and anti-actin (third panel) antibodies, as describe in “Materials and methods”. Immunofluorescent staining of control (Vo) and heparanase (Hepa) transfected cells is shown in the lower panels applying anti-heparanase monoclonal antibody (green). Nuclei counter staining is shown in red. Shown are representative results obtained by prostate carcinoma LNCaP cells. B. Control (Vo) and heparanase transfected (Hepa) prostate carcinoma LNCaP (I panels), epidermoid A431 (II panels), breast carcinoma T47 D (III panels), and melanoma MDA-MB-435 (IV panels) cell lysates were subjected to immunoblotting applying anti VEGF C (upper panels) and actin (lower panels) antibodies. VEGF C induction by heparanase in MDA-MB-435 cells is further demonstrated by RT-PCR (V panels) and immunofluorescent staining applying anti-VEGF C antibody (green). Nuclei counter staining is shown in red (C). D. siRNA experiments. LNCaP cells were left untreated as control (Con) or transfected with control (si-Con) or anti-heparanase (si-Hepa) siRNA constructs and VEGF C levels were evaluated by immunoblotting (upper panels) and RT-PCR (lower panels) analyses. Actin (second panel) and GAPDH (lower panel) were used as internal controls.