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Initially, we established that knockdown of myospheroid led to a significant decrease in cell speed (mean cell speed 1.54±0.11 µm.min−1) when compared to wild-type (mean cell speed 2.77±0.60 µm.min−1) in a similar manner as observed in the mys hemocytes (Fig. 1E).
With the addition of TGF-β, mean cell speed significantly decreased (−23%) at lower collagen concentrations (2.5 mg/ml), but increased or remained the same in denser matrices (4 and 6 mg/ml respectively).
Cells with a goodness-of-fit R2<0.5 were not included in the calculation of mean cell speed.
In fact, removing the non-motile cells from the analysis decreases the difference of mean cell speed under the level of statistical significance (Fig. 9F right panel).
Analysis of FLNabKD1 cells also revealed a significant increase in the percentage of non-motile cells (Figure 5G), but in this case the reduction in mean cell speed was not statistically significant (Figure 5H).
We found that knockdown of either isoform caused a ∼35% reduction in mean cell speed relative to control, indicating that both α-actinin-1 and α-actinin-4 contribute to glioma cell motility.
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Equating the experimental and simulated mean cell
Figure 3a shows time-dependent effects of 3 mM VPA on the mean-cell speed of L929 cells.
Interestingly, a time-response of the VPA-induced change in L929 mean-cell speed exhibited a biphasic response, with a significant reduction detectable already after ~20 min followed by a further decrease after 24-48 h.
50-200 cells from each well were tracked, and the data were used for the calculation of the mean-cell speed and the mean-squared cell displacement (< d >) as previously described [ 19].
The mean cell instantaneous speed is the mean of all instantaneous speeds of a cell.
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