Figure 5

Developmental process-specific depletion of essential proteins. (A) Meiosis-specific depletion of Cdc14. Yeast cells with P _CYC1 -GFP-TDegF-CDC14 were subjected to sporulation in the absence or presence of P _IME2 -GFP-pTEV⁺. Samples were taken at the indicated time points after induction of meiosis due to shift on sporulation medium (spo). The proteins (GFP-TDegF-Cdc14 and GFP-pTEV⁺ protease) were detected with antibodies directed against GFP; tubulin was used as loading control. (B) Depletion of Cdc14 or Cdc5 leads to a block during meiosis I. Yeast cells sporulated for 24 hours were stained with Hoechst 33342 to visualize the DNA. Bright field (DIC) and fluorescent (DAPI) images (maximum intensity projections) are shown. The genotypes of the strains are indicated in the figure. The CDC5-GFP-cODC1-TDegF cells contained a high copy plasmid with P _IME2 -GFP-pTEV⁺. Bar size, 2 μm.