Mutant TERT promoter displays active histone marks and distinct long-range interactions: A, cell lines that were used in the study with their origin and.

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Mutant TERT promoter displays active histone marks and distinct long-range interactions: A, cell lines that were used in the study with their origin and TERT promoter status. Mutant TERT promoter displays active histone marks and distinct long-range interactions: A, cell lines that were used in the study with their origin and TERT promoter status. WT refers to wild-type TERT promoter; WT-ALT refers to wild-type TERT promoter with alternate lengthening of telomeres. B, schematic view of TERT promoter. TSS represents the transcription start site. TERT promoter mutations were indicated as -146C>T and -124C>T. TERT promoter is shown in three parts including proximal promoter (up to -1 kb), distal promoter (-1 kb to -5 kb), and long-distance elements. C, ChIP was performed in A375 and BLM melanoma cells against histone marks (H3K4Me3 and H3K9Ac). D, ChIP was performed in primary melanocytes, Fadu, and PC3 cell lines against histone marks (H3K4Me3 and H3K9Ac). Graph shows qPCR analysis with % input obtained across various regions of TERT promoter with the distances indicated in boxes below the x axis. Error bars indicate the mean ± SD of the two independent experiments. E, 4C sequencing for long-range interactions of TERT promoter. Plot generated by r3CSeq of chromatin interactions of the TERT promoter for A375 and BLM cell lines. The top panel shows the RefSeq genes in chromosome 5. The line plot shows detected interactions 500 kb upstream and downstream of the TERT promoter. The x axis indicates the distance from TERT promoter which was shown as “0.” The y axis was the read counts for each interaction. Different shades of red and blue color dots indicate positive interactions for A375 and BLM cells (average of two replicates), respectively. Darker color indicates more significant interactions according to q values indicated in the legend. P values were calculated by two tailed Student t test method. Semih Can Akıncılar et al. Cancer Discov 2016;6:1276-1291 ©2016 by American Association for Cancer Research