Schematic representation of TERTp with the relative positions of cg11625005 (position 1,295,737 in hg19) to the TERTp mutations (position 1,295,228 and 1,295,250) and the transcription start site (TSS). A. Heat-map of methylation fraction (MF) in 31 CpG sites (top) in 44 samples (left). Yellow-marked CpG cg11625005 (position 1,295,737) is recognised by MSRE HgaI. Blue-marked CpG in 1,295,731 is recognised by MSRE AvaI. Black rectangle: MF at the cg11625005 measured either by NGS (clear squares, n = 44) and by ddPCR (patterned squares, n = 17; these samples were not included in the 44-sample batch subjected to NGS). B. TERT mRNA expression in 31 samples by qPCR analysed through the ΔΔCT method in Bio-Rad CFX manager software (version 3.1, Bio-Rad). C. TERTp mutations evaluated through ddPCR with commercial TERT C250T and C228T Mutation Assays in total 61 samples. D. Analysis of the chromatin accessibility in 8 cultured cell lines for TERT methylation region using GAPDH as a positive control.