Fig. 2.

TERT regulates miR500A by directly binding to its promoter region. (A) Schematic representation of the miR500 cluster according to the Ensembl database. Names are shortened to simplify. (B) Quantification of CLCN5 mRNA levels in TERT overexpression conditions by real‐time RT‐qPCR. (C) miR500A promoter activity in TERT overexpression conditions and (D) in TERT‐depleted HEK293 cells by luciferase reporter assay. (E) Determination of the promoter occupancy by amplification using a ChIP assay in hTERT‐SAOS 2 cells. The schemes represent the primers mapping to both negative (intron_GAPDH) and positive (TBE_cMyc) controls, and to the miR500 cluster. Each bar represents the mean ± SEM from triplicate samples. Graphs are representative (B, E) or the average (C, D) of three (N = 3) (B‐D) or two (N = 2) (E) independent experiments. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001 according to Student's t‐test (B, C) and ANOVA followed by Dunnett's multiple comparison test (D, E).