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Figure 2 DAbindinging to dead LL2 cells in vitro.
Tumour DAB4 binding was commensurate with treatment-induced cell death (Figure 5E), due in part to DAB4 binding to dead tumour cells (Figure 5F).
LL2 cells were treated with cisplatin for 48 h and (A) DAB4 binding or (B) Sal5 binding to dead cells was examined by flow cytometry.
Moreover, we suggest that vitronectin binding to dead cells may represent one of the mechanisms of vitronectin incorporation into the injured tissues.
Non-specific-binding can be the result of Fc receptor-binding, binding to dead cells, or by improper use of reagents.
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We have shown previously that one of these fluid-phase complement inhibitors, C4b-binding protein (C4BP) binds to dead cells and provides protection against excessive complement activation [16] [18].
This, and the loss of cell membrane integrity during cell death, make La accessible to DAB4 binding and means that DAB4 preferentially binds to dead tumour cells[10].
When cell viability under the optimized conditions was assayed by the production of calcein from calcein-AM (live cells) or PI binding to DNA (dead cells), the overwhelming majority of adipocytes remain viable compared to the dead cells as assayed using fluorescence detection of calcein and DNA bound propidium iodide (Figure 4).
In contrast, chemotherapy not only markedly increased the frequency of dead tumor cells (Figure 2A) but also produced a cumulative increase in DAB4 binding to each dead tumor cell (Figure 2B), and suggested that DNA-damaging drugs induced tumor cell expression of La in vivo.
RNA binding to the DEAD-domain then completes cleft closure to allow formation of an active ATPase site (Figure 6).
Studies in D. melanogaster showed that the HPat N-term sequence confers binding to the DEAD-box protein Me31B (Saccharomyces cerevisiae Dhh1 and human DDX6/RCK), whereas the Mid domain is necessary and sufficient for LSm1 7 binding (Haas et al, 2010).
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