FIGURE 4: SNF1 expression and Fkh protein abundance are decreased upon Ubc1 deletion.

(A) SNF1 and rRNA products from RT-PCR (26 cycles) were run on an agarose gel. RNA was isolated from isogenic WT and ubc1Δ strains that were grown to early log phase.

(B) Quantitation of three biological repeats of (A) is shown with mean, SEM, and t-tailed significance (Prism 6.0).

(C) The transcription factors Fkh1 and Fkh2 were endogenously TAP-tagged within the WT and ubc1Δ strains, and steady-state protein levels in 2% glucose (Fkh1-TAP left panel, 15 μg/lane, and Fkh2-TAP right panel 80 μg/lane) assessed by Western analysis. Clb2 is detected endogenously.

(D) Light microscopy (100x objective) of an early logarithmic asynchronous culture showed the proportion of large-budded cells, with corresponding flow cytometry identifying the relative population of cells with replicated DNA (2n) in ubc1Δ compared to WT strains.