Fig. 2

Netrin-1 co-exists with glioblastoma stem like cells in tumor stroma and infiltrative sprouts. a Colocalization of NTN1 and Jagged1 was analyzed by immunofluorescence staining of human GBM tissue. Representative area of the full image is surrounded with white box and presented as enlargement in lower panel. Scale bars represent 50 μm in full image and 20 μm in the enlargement. Merged image with all channels and separate channels of the enlargement are presented. NTN1 is marked with green, Jagged1 with red and nuclei with blue. b Colocalization of NTN1 and Notch2 was visualized by immunofluorescence staining of human GBM tissue similarly as in (a). NTN1 is marked with green, Notch2 with red and nuclei with blue. c Colocalization of NTN1 and Nestin was analyzed by immunofluorescence staining of human GBM tissue similarly as described in (a). NTN1 is marked with red, nestin with green and nuclei with blue. c Colocalization of NTN1 and CD133 was studied by immunofluorescence staining of human GBM tissue similarly as described in (a). NTN1 is marked with green, CD133 with red and nuclei with blue. e and f To study invasive front of human GBM tumor, tissue pieces were grown in 3D Matrigel ex vivo. The location of NTN1 and nestin (e) or NTN1 and Notch2 (f) was analyzed with immunofluorescence microscopy. The border of tumor tissue and invasive cells is implicated with dashed line. Enlargement of the boxed area is presented. In the enlargement white arrow points the direction of the migration. Green arrows point to NTN1 positive cells and red arrows Nestin positive cells in (e) and Notch2 positive cells in (f)