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By using two sequential selections based on LEU2 (growth on leucine-deficient media) and lacZ (blue color on X-gal plates) reporter gene activity, false-positive clones were reduced, resulting in a high proportion of cDNAs that were clearly true-positives (124 of 276 total cDNA hits tested from 11 separate cDNA library screening experiments).
P-PKB levels in the wild-type tissue neighboring with PTEN clones were reduced as compared to the tissue neighboring with control clones.
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However, the number of simultaneously competing clones is reduced due to the faster sweeps.
Interestingly, the rate at which Bu-1alow variants were generated by the rev1: BU-1AΔ G4 clones was reduced relative to the parental rev1 mutant, but was still well above the background level observed in wild-type clones (Fig 2E).
Third, the diameter of the branch in the region affected by Sal-expressing clones was reduced in the cells that expressed Sal and greater in adjacent regions that expressed vein.
Contigs with more than 10% Q-clones were reduced by running the DQer with a stricter cutoff by setting the value of DQer function to 10% and the step value to 5.
At higher stringencies (1e-14 to 1e-30), "contigsc" contigs were split and Q-clones were reduced, but the number of singletons increased drastically.
At higher stringencies (1e-20 to 1e-10), contigsc contigs were split and Q-clones were reduced, but the number of singletons increased drastically (Additional file 7: Figure S3).
Apical Dachs levels within the clone are reduced, and Dachs is enriched at the edge of the clone.
After additional months of treatment two small T315I positive clones emerged, carrying the D276G (2%) and the H396R (1%) respectively, while the F359C positive clone was reduced by half.
These data suggest that in the most chemoresistant samples, the leukaemic clone is reduced rather than eradicated, such that relapse is high in these cases.
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