Figure Legends
Figure 1. ALK protein immunocytochemistry on cell block sections of neuroblastoma.
A. Completely negative; B. 1+ granular cytoplasmic positivity; C. 2+ granular cytoplasmic positivity, D. 3+ strong diffuse ALK positivity which is cytoplasmic as well as membranous, E. Granular positivity seen inside the neuropil [Immunoperoxidase; o.m – A-Ex200], F. Dot plot of ALK H-scoring done in ascending value. Each dot represents a case. The dotted line marks the cut-off value of 100.
Figure 2. ALK gene mutated undifferentiated neuroblastoma in a 10-month-old. A. Cellular FNA smears with undifferentiated small blue round cells with high mitosis and karyorrhexis; B. Cell block immunocytochemistry showing ALK protein expression; C, FISH showing MYCN amplification; D, Screenshot from Integrated genomics viewer showing ALK gene mutation on locus chr2: 29443695 resulting in F1174L [A, May-Grünwald-Giemsa stain; B, immunoperoxidase stainx200; C, FISH Vysis LSI N-MYC (2p24) spectrum green/ CEP 2 spectrum orange FISH probe with DAPI counterstain; A x400, B x400, C x1000 o.m.]Figure 3. IDH1 gene mutated poorly differentiated neuroblastoma. A-B. Cellular smear and corresponding cell block showing rosetting of neuroblastic cells with abundant neuropil in the background; C. Cell block immunocytochemistry showing no ALK protein expression; D. FISH showing non-amplified MYCN; E. Screenshot from Integrated genomics viewer showing IDH1 exon 4 missense pathogenic mutation on locus chr2: 209113151 resulting in amino acid change R119Q. [A, May-Grünwald-Giemsa stain; B, Hematoxylin-Eosin stain; C, immunoperoxidase stainx200; D, FISH Vysis LSI N-MYC (2p24) spectrum green/ CEP 2 spectrum orange FISH probe with DAPI counterstain; A-C x200, D x1000 o.m.]
Figure 4. A. ALK protein expression in the three INRG subgroups. B-D. Kaplan-Meier curve showing relationship of overall survival in neuroblastoma with ALK (positive versus negative cases), combination of ALK and MYCN (4 subgroups) and combination of ALK and morphology (4 subgroups). Tick marks indicate censored individuals.
Supplementary Fig. 1. Cox Proportional Hazard Model and Forest plot showing Hazard ratios (HR) with 95% confidence interval.
Table 1 . Association between morphology and MYCNstatus parameters in ALK positive cases