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RLS was measured for individually selected virgin cells.
After their first division, mothers were removed and 2nd generation virgin cells kept for analysis.
This probably explains the overall difference in take rate (1 in 2,608 for virgin cells from Siwko and colleagues, and 1 in 20,636 for virgin cells from our data; see discussion above).
Briefly, virgin cells from exponentially growing cultures were placed on YPD plates using MSM Singer Micromanipulator (Singer Instruments, Roadwater, Somerset, UK).
For the 2nd round, single virgin cells from these colonies were subjected to replicative lifespan analysis, and 5th (from each young line) and last (from each old line) daughters were collected again, to obtain additional colonies.
Moreover, in this putative anamnestic response the immune cells detect antigenic differences in the cell populations of embryos of varying age, which are not as readily demonstrable when cytotoxicity is derived from virgin cells.
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These markers define the Immune Risk Phenotype (IRP) which is characterized by a depletion of CD95- virgin T cells and the expansion of CD28- T cells, resulting in an overall reduced T cell repertoire in old ages [28].
Virgin daughter cells were isolated and allowed to grow into mother cells while their corresponding daughters were microdissected using Zeiss Axioskop 40 dissection microscopes and manually counted until the mother cell could no longer divide.
It was found that several intrinsic material parameters, including V e, d i, W a − W c (the difference between the work functions of the anode and cathode), influence the programming time of the virgin Ag/GeS2/W cells.
For samples subjected to UV irradiation, all procedures were the same except that the virgin mother cells were UV irradiated and incubated for 1.5 h in the dark before counting the buds.
Normal MEC subsets, including MaSCs, LPs, and MLs, were sorted from WT C57/B6 adult virgin females; alveolar luminal cells (ALs) were sorted as YFP+ cells from Wap-Cre R26Y Wap-Cre R26Yid-gestation.
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