Fig. 6
The ROR2 receptor is a robust ALK+ ALCL marker. A. Heatmap representation of genes showing progressive upregulation: from activated T lymphocytes to NA cells in vitro, then from NA cells in vitro to in vivo models (a threshold of twofold increased expression was applied for each assessment) in the model dataset. B. Heatmap representation of the list of genes found in A in the patient dataset (n = 39). Log2 transformed expression matrix was used to generate representations. C. Western blot analysis of ROR2, NPM-ALK and phosphorylated NPM-ALK (P-NPM-ALK) in activated T lymphocytes, ALKIma1 cells, an ALK+ ALCL cell line (SUDHL1), patient PDX cells and an NPM-ALK overexpression model (NA-OE). NBS1 was used as a loading control. D. Western blot analysis of ROR2, NPM-ALK and phosphorylated NPM-ALK (P-NPM-ALK) in NA cells 16 days and 3 months post-transfection in activated T lymphocytes, ALKIma1 cells, and patient PDX cells. NBS1 was used as a loading control. E. Immunofluorescence analysis of ROR2 expression in ALKIma1 and PDX patient cells. Green: ROR2, red: actin, blue: DAPI. F. Histologic analysis of a CD4+ spleen tumor (corresponding to mouse #925 injected with NA4-E) and a CD8+ spleen and lymph node tumors (corresponding to mouse #791 injected with NA5-ECD8): anti-ROR2 IHC. The negative control spleen (CTL) was obtained from a wild type NSG mouse. G. Histologic analysis of ALCL patient tumors: anti-ROR2 IHC. Left: NPM-ALK+ ALCL tumors (P1, P11, P13), middle: ALK+ ALCL tumors (other fusion partners) (P4, P23 and P14); right: ALK- ALCL tumors (P26 and P27); ROR2 negative tumor (P25). See also Table 1 for all sample data