Figure 6.

orb6⁺ gene product intracellular localization in the cell cycle. (a, b, e, and f) orb6∷ura4⁺ ade- leu1–32 ura4-D18 cells containing integrated HA-tagged orb6⁺ were grown exponentially at 32°C in the absence of thiamine, then fixed and stained with an antibody recognizing the HA tag (a and b) and with 4′,6-diamidino-2-phenylindole (e and f). (g) Control staining of orb6∷ura4⁺ ade- leu1–32 ura4-D18 cells containing integrated untagged orb6⁺. (c) cdc25–22 leu1–32 Rep3X orb6 HA cells were grown exponentially at 25°C in the presence of thiamine for at least eight generations; thiamine was washed away and cells were grown in the absence of thiamine at 25°C for 16 hr, then shifted at 36°C for 4 hr. (d) orb2–34 leu1–32 pRep3X orb6 HA at 32°C; cells were stained 14 hr after thiamine depletion, which is about 2 hr after derepression of the nmt1 promoter. (h) orb2–34 leu1–32 stained for actin at 32°C. (Bar = 10 μm.)