Fig. 5. Localization of Pub2p. (A) Localization of Pub1-GFP and Pub2-GFP. pREP81-based plasmids carrying GFP, Pub1-GFP or Pub2-GFP fusion constructs were introduced into the wild-type strain MM72-11C. After a 12 hour incubation on thiamine-free EMM2, cells were observed under a fluorescence microscope. (a) GFP, (b) Pub1-GFP and (c) Pub2-GFP. Scale bar, 10 µm. (B) Fractionation of cell lysates. Cell lysates were prepared from the Pub2-HA integrant (KKT87). P13 is a precipitate after centrifugation at 13,000 g, and P100 and S100 fractions are the sediment and the supernatant, respectively, after the second centrifugation at 100,000 g. Pub2-HA was detected by western blotting with anti-HA antibody 3F10.