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. 2014 Aug 1;4:5924. doi: 10.1038/srep05924

Figure 4. Podoplanin protein expression was determined immunohistochemically in paraffin-embedded tumor specimens.

Figure 4

Histologic sections 4 µm thick were deparaffinized in xylene and rehydrated, then autoclaved in citrate buffer (pH 6.0) for 20 min. Sections were incubated with 5 μg/ml of LpMab-2 (a-e) or LpMab-7 (f–j) overnight at 4°C with subsequent treatment using an LSAB kit. Color was developed using 3, 3-diaminobenzidine tetrahydrochloride (DAB) for 10 min and was counterstained with hematoxylin. (a, f) Esophageal squamous cell carcinomas include both cancer cells (upper) and lymphatic endothelial cells (lower). Cancer cells were stained with both LpMab-7 (f) and LpMab-2 (a), whereas lymphatic endothelial cells were stained with LpMab-7 (f), not with LpMab-2 (a). (b, g) Seminoma includes both cancer cells (left) and lymphatic endothelial cells (right, upper). Cancer cells were stained with both LpMab-7 (g) and LpMab-2 (b), whereas lymphatic endothelial cells were stained with LpMab-7 (g), not with LpMab-2 (b). (c, h) Glioblastomas were stained with both LpMab-7 (h) and LpMab-2 (c). (d, i) Esophageal lymphatic endothelial cells were stained with LpMab-7 (i), not with LpMab-2 (d). Lung type I alveolar cells were stained with LpMab-7 (j), not with LpMab-2 (e). Arrows indicate lymphatic endothelial cells (f, g, i).