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. 2018 Mar 8;8(5):1579–1592. doi: 10.1534/g3.118.200161

Figure 5.

Figure 5

Vacuolar acidity is controlled by Fkh1 or Fkh2. (A) Native chromatin immunoprecipitation (ChIP) of select VMA promoters in cells expressing the TAP epitope fused to the C-terminus of FKH1 or FKH2 (Postnikoff et al. 2012). Genomic DNA in TAP immunoprecipitates was amplified using primers to 5′ upstream regions. See Figure S4 for FKH consensus binding sites within VMA promoters. (B) Mutants harboring deletions of either or both FKH genes were grown overnight in YPD, then stained with cDCFDA and imaged in parallel. Deleting both FKH genes induced nutrient-independent pseudohyphal growth (Zhu et al. 2000; Hollenhorst et al. 2000). (C) qPCR analyses of VMA mRNA expression ratios in fkh1Δ fkh2Δ mutants relative to WT cells. Mean ± SD shown. (D) RT-PCR analyses of FKH mRNA in young and old cells enriched by MEP. Expression ratio of FKH genes in old cells relative to young cells. Mean ± SD shown.