Figure 5. The combination of radicicol and hygromycin B extincts karyotypically heterogeneous cell population.

(A-E) Chr XV trisomy was able to escape growth inhibition by hygromycin B through loss of the gained Chr XV. (A) The growth (represented by OD reading on a Tecan reader) of both the euploid control and the trisomy XV strain was monitored in media containing 50 μg/ml hygromycin B. (B) The additional copy of Chr XV was lost in hygromycin B culture but not in YPD culture, as shown by the heat map of karyotyping result of the final culture. (C) Karyotypes of 6 single colonies from the trisomy XV culture in YPD or hygromycin are shown, three of which were re-tested for growth in the presence of hygromycin B (D) or radicicol (E). Note radicicol sensitivity was re-established in all three adapted colonies from the trisomy XV culture in hygromycin, whereas isolates from the YPD culture remained radicicol resistant. (F-H) Combination of hygromycin B and radicicol effectively eradicates the radicicol-preselected aneuploid population. (F) Growth curves (as OD600 measured in Tecan) of the diploid control strain under conditions as indicated. Note that 50 μg/ml hygromycin B alone had milder growth suppression compared to 100 μg/ml radicicol. (G) Growth curves of 3 populations preselected independently in the presence of radicicol (Figure S3E) under indicated conditions. (H) Growth curves of the same 3 populations as in (b) under indicated conditions where each drug was combined with 50 μg/ml radicicol. Each data point in B and C shows the mean and SEM from 3 experiments.

See also Figure S6.