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. Author manuscript; available in PMC: 2016 Mar 9.
Published in final edited form as: Cell Rep. 2016 Jan 28;14(5):1010–1017. doi: 10.1016/j.celrep.2016.01.005

Figure 4. Condensin and Hmo1 mediate the transcriptional position effect in the rDNA array.

Figure 4

(A) Repeat-specific URA3 expression was measured in nutrient-depleted ycs4-2 mutants at 23°C by qRT-PCR. Signals were normalized to UBC6 and repeat 29. Error bars indicate s.d. of 3 independent experiments, each analyzed in triplicate. (B) Plate growth assays of wild-type, ycs4-2 and ycg1-2 cells carrying repeat-specific reporter cassettes. 3.5-fold serial dilutions were plated onto synthetic glycerol medium supplemented with uracil (complete) or lacking uracil (-ura) and grown at 23°C. Arrows highlight the dilutions with the most obvious position-dependent differences in colony size. (C) Repeat-specific URA3 gene expression was determined in nutrient-depleted hmo1Δ cells by qRT-PCR. Values are normalized against UBC6 and repeat 49 for each condition. Because of the slow growth of hmo1Δ mutants, cells were harvested after 24 and 48 hrs of nutrient depletion. 48 hrs of nutrient depletion in hmo1Δ mutants roughly corresponds to 24 hrs of nutrient depletion in wild type, as determined by growth curves. Error bars as in (A). Significance in (A) and (C): Welch two-sample t-test. (D) Plate growth assays of hmo1Δ mutants carrying repeat-specific reporter cassettes. Analysis as in (B) but cells were grown at 30°C. See also Figure S4.