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In the leukemic cell data presented, exposure to ganetespib resulted in G1 and G2/M arrest, in part through the degradation of Cdk1 and atypical accumulation of cyclins A1 and B1.
The single cell data presented in Figure 2B did not include measurement of both transcripts within each individual cell and could not determine whether the small number of cells showing increased IFNB1 and DDX58 levels in the presence of blocking antibodies were the same subset of cells.
Consistent with the whole cell data presented above, after RNAPII inhibition, not only were CENP-A signals reduced on chromatin fibers, HJURP was almost completely lost.
Live cell data presented here provide a framework to explain how extracellular factors acting via small GTPases of the secretory pathway may regulate connexin transport to the cell surface.
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Mutation frequency was calculated as the number of CanR mutants per 10 surviving cells (data presented in Figs 3B and 4A and B).
The analysis revealed that the spindle orientation of p600 RNAi-electroporated RGCs deviates by ∼29% from the mainly vertical cleavage planes of control cells (data presented as mean ± SD: control: 73.1±16.9°, n = 61 cells; RNAi #1: 54.2±25.5°, n = 44 cells; U = 644, p<0.000005; RNAi #2: 52.3±26.9, n = 34 cells; U = 555, p<0.0001 by one-tailed Mann–Whitney U test) (Fig. 2B).
DNA copy number for each set was quantified for 100 cells and data presented per cell.
The cell viability data presented thus indicate the percentage of cells that were multiplied over a three days period and not the percentage of cells that survived a toxic insult.
Cell survival data presented in Figure 2 show that two distinct groups of cell lines can be distinguished: TMZ sensitive and TMZ resistant.
The cell line data presented here may offer an explanation for this in that cell lines with Syndecan predominant HSPG pattern (or perhaps, as described for survival in other cancers, cells that have lost CD44v3 expression) become more platinum resistant when chronically exposed to DcR3 while the cell line with CD44v3 as a more dominant HSPG become more platinum sensitive.
This hypothesis is corroborated by our cell cycle data presented here.
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