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A reduction of FLIP levels was associated with an increased sensitivity to TRAIL-mediated cell death (Zhang et al, 2000; Hernandez et al, 2001).
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A reduction of FLIP-L was recorded in several experimental conditions, especially under the combined treatment.
In contrast, H. pylori significantly enhanced the recruitment of caspase-8 into the TRAIL DISC through reduction of the FLIP assembly and therefore augmented the activation of caspase-8.
AR-42 also sensitizes CLL cells to TNF-Related Apoptosis Inducing Ligand (TRAIL), potentially through reduction of c-FLIP.
This is potentially due to reduction of c-FLIP protein, an effect we previously reported in CLL cells using romidepsin [10].
Since the concentrations of MG-132 used here are very low, it is possible that reduction of c-FLIP by MG-132 is independent of its ability of inhibiting proteasome activity.
A study in colon cancer cell lines [46] showed that the DAC inhibitor sodium butyrate also caused substantial decrease in c-FLIP protein concurrent with TRAIL sensitization, although similar studies in several hematological cell lines using sodium butyrate and vorinostat demonstrated TRAIL sensitization without reduction of c-FLIP [34].
Analysis of mRNA expression confirmed reduction of c-FLIP mRNA expression following transfection with siFGFR4, and a reduction in Bcl-2 expression was also observed at 48 h.
The increased susceptibility in K8-null liver was reported to be due to a drastic reduction of c-Flip protein and associated with inhibition of phosphorylation/activation of p44/42 MAPKs (Gilbert et al., 2004).
In an other report it was shown that the reduction of c-FLIP, Bcl-xL, Bcl-2, and XIAP expression induced by subtoxic doses of the HDACI LAQ824 was independent on proteasome or caspases activity [ 26].
Although TRAIL-R1 and R2 were minimally affected by the treatment at the mRNA level (data not shown), we did observed a reduction of c-FLIP expression (Supplementary Figure 3).
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