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. 2015 Dec 14;10(12):e0144972. doi: 10.1371/journal.pone.0144972

Fig 2. A new IPL1 conditional allele.

Fig 2

(A-B) Cells were grown at 25°C in SC medium without methionine (SC-MET), arrested in G1 with 5 μg/ml α-factor, and released at 37°C into SC medium containing (+MET) or not (-MET) 8 mM methionine and without pheromone. (A) Western blots show Ipl1-3HA levels at the indicated time points and in a cycling culture of pMET3-Ub-DHFR-IPL1-3HA (F1595) cells. Pgk1 was used as loading control. (B) Percentages of wild type (F955), ipl1-321 (F323), or pMET3-Ub-DHFR-IPL1 (F1517) cells displaying correct (white bars) and incorrect (black bars) segregation of CrIV-GFP in anaphase. Error bars indicate SD (n = 3). (C) 10-fold serial dilutions of wild type (F496), ipl1-321 (F323), or pMET3-Ub-DHFR-IPL1 (F1124) liquid cultures grown in SC-MET at 25°C were spotted on SC-MET and SC+MET plates at the indicated temperatures to analyze cell viability.