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