Table 4. Mutations defined as pathogenic in previous cohorts.

Publication	Nr of POLE mutated tumors	Sequencing method	Mutations included in the “POLE mutated” group	Definition of pathogenic mutation
Tomlinson et al 2013	14	Sanger sequencing (codons 268–471)	P286R, S297P, V411L, A456P, A275V	Predictive and functional analysis
McAlpine et al 2015	39	Sanger sequencing exons 9 to 14	P286R, S297P, V411L, A456P, M295R, F367S/C, P436R, L424P, P441L, F367L, E396G
Ngeow et al 2016	12	Nextgeneration sequencing and sanger sequencing to confirm	P286R, V411L, A456P, S459F A465F, M444K, S459P	In silico testing of mutations to define pathogenity
Billingsley et al 2015 and Goodfellow et al 2017	40/39	Sanger sequencing (residues 268–471)	P286R, S297P, V411L, A456P, P436R, A465F, A426V	assessed using mutation assessment prediction programs
Bosse et al 2015	63	Sanger sequencing exon 9 and 13 (PORTEC)	P286R, S297P, V411L, M299V, S297T	defined as pathogenic POLE proofreading mutations as variants absent from public germline sequence databases and previously confirmed as somatic variants associated with tumor ultramutation
Köbel et al 2014	8	Sanger sequencing exon 9–13	P286R, V411L, T278M, S297P	all mutation positive samples

References [10,40–44]