Ssu72 is required for telomeric C‐strand stability Ssu72 is required for telomeric C‐strand stability ssu72Δ telomeres present longer G‐rich overhangs than wt and rif1∆. In‐gel hybridization in native and denaturing conditions was labeled with a radiolabeled C‐rich telomere probe and quantified for ssDNA at the telomeres. n = 3; *P ≤ 0.05 based on a two‐tailed Student's t‐test to control sample. Error bars represent standard error of the mean (SEM).RPA (Rad11‐GFP) is enriched at ssu72∆ telomeres. Co‐localization of Rad11‐GFP with Taz1‐mCherry, used as a telomere marker, was performed in wt and ssu72∆ cells; n = 3; *P ≤ 0.05 based on a two‐tailed Student's t‐test to control sample. Error bars represent standard error of the mean (SEM). More than 1,000 cells were analyzed for each genotypessu72+ controls telomere length independently of rif1+. Epistasis analysis of telomere length of ssu72∆ and ssu72‐C13S (catalytically inactive mutant) with rif1∆ was performed by Southern blotting of ApaI‐digested DNA using a telomeric probe.ssu72+ and stn1+ regulate telomere length in the same genetic pathway. Epistasis analysis of ssu72∆ and stn1‐75 performed by Southern blotting of ApaI‐digested DNA using a telomeric probe. Two independently generated ssu72∆ stn1‐75 double mutants are shown. Jose Miguel Escandell et al. EMBO J. 2019;38:e100476 © as stated in the article, figure or figure legend