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Page 10 of 14 Schwertheim et al. Hepatoma Res 2020;6:41 I http://dx.doi.org/10.20517/2394-5079.2020.23

Figure 5. Kaplan-Meier survival studies. Disease specific OS was analyzed in 69 valid hepatocellular carcinoma (HCC) samples in
relation to the presence of KDM2A immunopositivity in intranuclear inclusions (A) and in the cytoplasm (B). Additionally, recurrence-
free survival was examined in 47 valid HCC samples in relation to the presence of KDM2A immunopositivity in the cytoplasm of tumor
2
(A) and in adjacent normal tissue (B). We counted inclusions in a standardized area of 23.5 mm (ten serial sections of each case). Only
cases with evaluable material on all ten serial sections were included. Cases containing at least one membrane-bound nuclear inclusion
were considered positive; P ≤ 0.05 was defined as statistically significant

Figure 6. Association between KDM2A immunopositivity and intranuclear inclusions (NI) in hepatocellular carcinoma. The diagram
depicts a significant association between positive KDM2A immunoreactivity within NI and the occurrence of NI. ***P ≤ 0.001

DISCUSSION
In the present study, we have demonstrated accumulation of β-catenin and proteins associated with the Wnt/
beta-Catenin pathway in NI, partly together with autophagy-associated proteins in the same inclusion. The
presence of inclusions with KDM2A immunopositivity correlated significantly with overall survival in HCC.
We found positive immunostaining for β-catenin, glutamin syntethase and KDM2A within NI [Figure 1]. In
addition, other immunohistochemical studies have reported strong immunopositivity for β-catenin within