Figure 4

Protein expression validation of ErbB3. A. Immunohistochemistry (IHC) assessment of case 2 showing ErbB3 expression in the mature ganglion cells, and Sox10 expression in both ganglion and schwannian cells. Tumour tissue sections were stained using horseradish peroxidase (HRP) secondary antibodies. The right panel shows the Sox10 and ErbB3 expression results from all IHC assessments performed, y = yes, n = no expression. B. Colocalization study of ErbB3 and Sox10, showing ErbB3 expression mainly in ganglion cells. Tumour tissue sections were simultaneously fluorescently stained using anti-goat Alexa Fluor 594 (Sox10, red) and anti-mouse Alexa Fluor 488 (ErbB3, green), and Dapi (DNA, blue). C. High protein levels in GN tumours shown by Western blot of ErbB3. Primary antibodies used in studies: Sox10 ([N-20], Santa Cruz Biotechnology), and ErbB-3 ([RTJ2], Abcam), which binds to both phosphorylated and unphosphorylated forms. Case 6 was also included in the microarray analysis (NBS1, data set 2). Clinical data: NB = neuroblastoma, GNB = ganglioneuroblastoma, GN = ganglioneuroma; INSS stage [2]; INRGSS stage [3]: L = Localised, L1/L2 (INSS 1, 2, 3); M = Metastatic (INSS 4); Outcome: NED = No evidence of disease (white); AWD = Alive with disease (grey); DOD = Dead of disease (black); MNA (MYCN-amplification), black = yes, white = no, na = not applicable.