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. 2005 Dec 19;171(6):967–979. doi: 10.1083/jcb.200504104

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Copyright © 2005, The Rockefeller University Press

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Figure 10. — Interallelic complementation of the inviability of pik1Δ cells by Pik1-CCAAX and Pik1(Δ10-192). (A) Heterozygous pik1Δ::KanMX4/PIK1 diploid (YTS68) cotransformed with either pTS2 and pTS4 expressing mycPik1(Δ10-192) and mycPik1-CCAAX (left) or pTS3 and pTS4 both expressing mycPik1-CCAAX (right) was sporulated and the resulting tetrads were dissected and germinated on galactose medium. Spore clones (A–D) from 10 representative tetrads (1–10) are shown. (B) Two representative G418-resistant colonies of opposite mating type were picked, propagated on galactose (Gal) or then shifted to glucose medium (Glc), and samples of the cultures were lysed and analyzed by SDS-PAGE and immunoblotting with αMyc mAb 9E10. As markers, samples of lysates from cells expressing only mycPik1(Δ10-192) (lane 5) or mycPik1-CCAAX (lane 6) were examined. (C) Tetrad analysis, as in A, was conducted with YTS68 coexpressing either mycPik1-CCAAX and mycPik1(Δ10-192 D918A) (left) or mycPik1(D918A)-CCAAX and mycPik1(Δ10-192) (right).