Figure 3.

Dependence of DNA Replication and Spindle Formation on CYCB1 and the APC.

(A) and (B) Cells were blocked by nitrogen deprivation and released at 33°C.

(A) DNA content analyzed by flow cytometry. Wild-type cells undergo multiple rounds of DNA synthesis and mitosis within the mother cell wall, producing cell clusters with 2, 4, 8, or 16C DNA content, before hatching to produce small 1C newborn cells. The apparent DNA content of these cells compared with predivision 1C cells is shifted to the left (Tulin and Cross, 2014).

(B) From a separate experiment, cells at 13.5 h after release were processed for anti-α-tubulin immunofluorescence and DNA localization by SYTOX staining. DIC, differential interference contrast image. Bars = 10 μm. Quantification is shown in Supplemental Table 3.