(A, B) SPB inheritance in wild-type (F2071) and cdc5-as1 (F2741) cells expressing Spc42-RFP and Spc110-GFP. (A) Representative images of Spc42-RFP (red) and Spc110-GFP (green) localization in cells exhibiting wild-type and reversed (old SPB retained in the mother cell) SPB distribution patterns. Also shown are nuclear morphology (DAPI, blue), bright-field (BF), and merged images. (B) The percentage of cells displaying wild-type (white bars) or reversed (black bars) SPB inheritance, based on Spc42-RFP distribution. (C–F) Spc72-GFP distribution and SPB inheritance in wild-type (F3712) and cdc5-as1 (F3707) cells also expressing Spc42-RFP. (C) Representative images showing coincidental or opposite Spc72-GFP (green) and Spc42-RFP (red) distribution, distinguishing in the former between cells exhibiting wild-type (main accumulation in the bud) and reversed (preferential retention in the mother cell) Spc72 localization. Also shown are nuclear morphology (DAPI, blue), BF, and merged images. (D–F) The percentages of cells displaying wild-type (white bars) or reversed (black bars) Spc72-GFP localization (D), wild-type (white bars) or reversed (black bars) SPB inheritance (based on Spc42-RFP distribution) (E), and coincidental (white bars) or opposite (black bars) distribution of both tagged proteins (F). (G) Spc72-GFP distribution in wild-type (F3696), cdc5-as1 (F3699), cdc5-2 (F4439), cdc5-77 (F4705), and cdc5-16 (F4699) cells, and localization of Spc72-AA-GFP mutant protein in a wild-type background (F4606). (H, I) Western blot analyses in asynchronous cultures of wild-type (F3696) or cdc5-as1 (F3699) cells expressing Spc72-GFP, and of wild-type cells expressing the Spc72-AA-GFP mutant (F4606) or untagged Spc72 (F496, No tag). (H) The experiment was performed thrice (n = 3); shown is a representative image displaying the levels of GFP-tagged protein. Pgk1 levels were used as the loading control. (I) Intensity plot profile of the lanes from the blot shown in (H). (J) SPB inheritance in wild-type (F3702) and cdc5-as1 (F3705) cells expressing Spc72-GFP, and in wild-type cells expressing Spc72-AA-GFP (F4608), all simultaneously carrying Spc110-dsRed. Shown is the percentage of wild-type (white bars) or reversed (black bars) SPB inheritance, based on Spc110-dsRed distribution. (B, D, E, F, G, J) Final data are the average of three experiments (n = 3; 100 cells each). Error bars represent the SD. Statistical significance according to the Newman-Keuls multiple comparison test is indicated.