Fig. 6.

xRRM domain of Pof8 is important for TER1-Pof8 interaction and accumulation of TER1 RNA. a Southern blot analysis showed that xRRM domain mutants show telomere shortening comparable to pof8∆. Genomic DNA samples from indicated strains were prepared after extensive restreaks on YES plates prior to analysis to achieve terminal telomere length. b TER1-Pof8 interaction was essentially eliminated in xRRM domain mutants of Pof8. The plot shows fold enrichment of TER1 RNA recovered after IP compared to no tag control strain. Error bars correspond to SEM from at least 6 independent experiments. c The xRRM domain mutants failed to accumulate telomerase RNA. TER1 RNA expression levels were normalized to his1⁺ mRNA expression. Error bars correspond to SEM from at least 5 independent experiments. d, e Quantitative real-time PCR ChIP analysis to monitor association with telomeres for (d) Pof8 and (e) Trt1 in indicated genetic backgrounds. Error bars correspond to SEM from at least 12 (d) and 8 (e) independent experiments. Raw data and statistical analysis are available in Supplementary Data 1. Expression levels of (d) Pof8 and (e) Trt1 used in ChIP assays were monitored by western blot analysis. Anti-Cdc2 blots served as loading control. Molecular weight (kDa) of size markers are also indicated