Supplemental Material

This article contains the following supporting material:

• Figure S1 - Localization of Kar9p-3GFP on cMTs. Examples of Kar9p-3GFP localization in wild-type (yRM3886), bik1Δ(yRM4261), and kip2Δ (yRM4371) cells expressing Tub1p-CFP labeled MTs. Arrowheads point to the Kar9p dot at the plus end of the cMT in kip2Δ cells. Cells were grown in SC media -TRP at 30°C.
• Figure S2 - Bik1p localization at cMT plus ends and SPBs. (A) The plus end localization of Bik1p does not require Kar9p. Bik1p-3GFP localization in wild-type (yRM3657) and kar9Δ (yRM3659) cells with Tub1p-CFP labeled MTs. Bik1p-3GFP is enriched at SPBs and the plus ends of MTs, and can also be seen along the length of MTs. Deletion of KAR9 did not disrupt the plus end localization of Bik1p-3GFP as compared to wild type. Bik1p-3GFP was observed at the plus ends of cMTs in 99% of pre-anaphase kar9Δ cells (n=157) and 94% of pre-anaphase wild-type cells (n=152). (B) Examples of Bik1p-3GFP localization in cells containing Spc110p-DsRed labeled SPBs (yRM4568). In cells with bipolar spindles Bik1p localizes to regions associated with both SPBs (n=150).
• Figure S3 - Bik1p functions in the restriction of Kar9p to one SPB. Examples of Kar9p-3GFP localization in wild-type (yRM4357) and bik1Δ (yRM4358) cells containing SPBs labeled with Spc110p-CFP. Composites are shown of DIC with GFP and CFP single images. In both strains Kar9p localizes to the unduplicated SPB in G1 and early S-phase, prior to SPB duplication. After SPB duplication, Kar9p is restricted to one pole in wild type, but localizes to both poles in bik1Δ.
• Figure S4 - Band intensity profiles for Kar9p-tap species. Intensity values were measured from Figure 5B using the plot profile feature of NIH Image version 1.63. These were then plotted using Microsoft Excel.
• Table S1
• Table S2