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To investigate upstream regulators of GSK3ß, quiescent PASMCs were stimulated with PDGF-BB (60 ng/ml) for 6 (Figure S3) and 24 hrs (Figure 4) or 10% FCS (Figure 5) for 24 hrs.
In this perspective it is important to investigate "upstream" influences on more proximal determinants of health [ 13].
The fact that we investigate upstream sequences for histone modifications might lead to under-estimate a putative 3metH3K9 modification in the sequences investigated for DNA methylation, and this results that RELB could be even more silenced than we proposed.
Overall, it will be important in future studies to investigate upstream activators of Sema3 expression and mechanisms that ultimately distinguish the morphological differentiation of type I and type II SGNs.
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Second, we investigated upstream pathways by measuring ERK MAP kinase activity and downstream pathways by determining the levels of the putative targets MMP-1 and MMP-7.
Furthermore, utilizing the IPA tool, we also investigated upstream regulatory molecules that are responsible for identified pathways and altered gene expression in stage II CRC.
The heat map of weight values of all motif pairs is provided in Additional file 3. Table 4 presents the top ten motif pairs with the highest weight values and with occurrence of more than ten in all the investigated upstream sequences.
To further investigate the upstream inhibition of mTOR by EMB, we investigated the phosphorylation levels of PI3K and Akt.
To investigate the upstream regulation of BB we combined a promoter deletion analysis with a phylogenetic footprinting approach.
To further investigate the upstream inhibition of MTOR by Mono-Pt, we examined AKT1 phosphorylation at Ser473 and Thr308.
As autophagy seemed to be differentially activated under normoxia and hypoxia, mTOR activity was studied to investigate the upstream signaling responsible for autophagy activation.
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