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. 2018 Nov 15;19(1):foy120. doi: 10.1093/femsyr/foy120

Figure 2.

Figure 2.

The t-Utps associate with rDNA sequences even outside of the rDNA repeats. (A) Semi-quantitative ChIPs of a subunit of RNA polymerase I (Rpa190), core box C/D snoRNP proteins (Nop1 and Nop5), core box H/ACA snoRNP proteins (Gar1), and t-Utps (Utp9, Utp15, Utp17/Nan1) tagged with HA in YPH499 were carried out. Probes used were to the rDNA promoter (primers –200 and 3΄ start), the 5΄ETS (primers +300 and oligo x) and the 25S rDNA (primers 5΄25S and oligo y; Table 1 for oligos). Lanes are marked B for beads alone in the immunoprecipitation; HA-indicates immunoprecipitation with the anti-HA antibody. (B) Schematic of the HOT1-his4 reporter from the K3207 strain. (C) Quantitative ChIP of HA-tagged Utp8 and Nop1 in K3207 using 5΄ start and 3΄ HIS4 primers that can only amplify the HOT1 sequence at the his4 locus but not at the rDNA. The signal was normalized to ACT1 and the standard deviation is indicated on the graph.