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For investigation of mode of action for compound 14 which displayed the most strong dual topoisomerase I and II inhibitory activity and antiproliferative activity against HCT15 cell, we performed cleavable complex assay, band depletion assay, comet assay, and competitive EtBr displacement assay.
To orient the HEV-VLP into the crystal unit cell, we performed a self-rotation search using the program GLRF (Tong and Rossmann 1997) as described below.
In order to determine whether peanut root tip cells underwent PCD during Al stress followed by the similar mitochondrial route as animal cell, we performed the immunoblotting detection of cytochrome c in mitochondria and cytosol.
To conclusively determine if the proliferative cell was indeed an endothelial precursor cell, we performed co-staining for phospho-histone H3 (p-H3) and GFP protein in Tg(fli1a: EGFP) 10 som section (Fig. 2K).
To eliminate the possibility that our subcellularly targeted GCaMP2 fusion proteins significantly perturbed the calcium channel or calcium handling machinery of the cell we performed additional experiments in which we simultaneously measured responses from the GECIs and from X-Rhod-5F, loaded through the patch pipette.
To ensure the siRNA treatments were not having adverse effects on the cell, we performed cell viability assays and found no difference in the viability of cells treated with control pool siRNA when compared to spectrin, adducin or p4.1 siRNA treated cells (Figure S11).
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To assess the proliferation and clonogenic potential of sorted cells, we performed a limiting dilution assay.
To better understand how ecdysone exerts its tumour-suppressor function in ph505 cells, we performed transcriptome analyses by mRNA sequencing.
To validate the Nef:MHC-I interaction in cells, we performed a bimolecular fluorescence complementation (BiFC) assay.
To quantitatively evaluate how the BM tissue landscape relates to these stromal cells we performed detailed spatial analyses.
To determine whether this loss in detection is due to rapid degradation or exit from the cells, we performed a cell-based sandwich ELISA (Supplementary Fig. S10).
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