Fig 2: Trans-differentiation of chromaffin cells in culture. Double-immunostaining shows that while TH (A) and TUBB3 (B) were maintained during the 6d culture period, the chromaffin cell-specific proteins PNMT, DLK1, and SGII are lost or decreased (E,F,G). Increased ISLET1 expression does not reach significance (C,H). PERIPHERIN is expressed by a very low number of cells both at 3d and 6d in culture (D,H). The results are quantified in (H). Significant differences marked by * and *** indicate p-values < 0.05 and <0.001, respectively. Magnification bar, 60 µm.

Fig 3: Characterization of mouse sympathetic neuroblasts. Dissociated E14.5 sympathetic ganglion cells were plated on porn/laminin-coated dishes in serum-free culture medium and analyzed after cell attachment. Double-immunostaining showed that all PHOX2B-positive cells co-express the noradrenergic markers TH (A) and VMAT1 (B) but not the chromaffin cell-specific proteins PNMT (C), DLK1 (D), and SGII (E). ISLET1 (F), TUBB3 (G), and PERIPHERIN (H) expression are shown as examples of neuron-specific proteins. Magnification bar, 60 µm.

Fig 4: Immunohistochemical staining of the micro-pheochromocytoma with (a) tyrosine hydroxylase (TH), (b) aromatic L-amino acid decarboxylase (AADC), (c) dopamine ß-hydroxylase (DBH), and (d) phenylethanolamine N-methyltransferase (PNMT) antibodies (×100). The primary antibodies were mouse monoclonal antibodies: anti-TH, F-11, sc-25269; anti-DDC, 8E8, sc-293287; and anti-PNMT, C-7, sc-393995, from Santa Cruz Biotechnology, Dallas, USA, and the anti-DBH goat polyclonal antibody, ab189991, from Abcam plc, Cambridge, UK. Dashed lines show boundaries between the normal adrenal medulla (m) and micro-pheochromocytoma (p). The letter c indicates the adrenal cortex.