Figure EV2. AGO1 binds to terc‐sRNA and to TERC, but AGO1 depletion does not affect telomerase activity neither in wild‐type nor in AGO2KO background.

• A
  Coverage of terc‐sRNA, as assessed by sRNA‐Seq of nuclear AGO1‐IP from HeLaS3 cells.
• B
  RIP assay was performed from HeLaS3 cell extract using anti‐AGO1 antibody or IgG, as negative control. Immunoprecipitation was verified by Western blot.
• C, D
  TERC RNA (n = 3 experimental replicates) and terc‐sRNA (n = 5 experimental replicates) enrichment in AGO1 RIP as compared to IgG RIP was assessed by RT–qPCR. 7SK RNA was used for normalization.
• E
  Telomerase activity was detected by TRAP in two HeLaS3 clones isolated following transduction with LentiCRISPR v2 control vector (HeLaS3 WT CRISP neg), AGO2KO cells following transduction with LentiCRISPR v2 (AGO2KO CRISPR neg), two AGO1/AGO2 double KO clones isolated from AGO2KO cells following transduction with LentiAGO1CRISPR v2 (AGO1/AGO2dKO), and two AGO1 KO clones isolated from HeLaS3 cells following transduction with LentiAGO1CRISPR v2 (AGO1KO). Quantitative analysis of telomerase activity was plotted (n = 3 experimental replicates).

Data information: Data are expressed as mean ± SEM. *P ≤ 0.05; **P ≤ 0.01; ns = not significant (Student's t‐test).