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To determine whether B cells contribute to CD4 T cell activation in the context of active disease, we evaluated the activation of both CD4 T cells and B cells by Leishmania antigens in PBMC cultures.
Using an antibody that specifically recognizes an epitope overlapping the nuclear localization signal of the p65 subunit and, thus, selectively binding to the activated form of p65 (indicated as p65*), we evaluated the activation of NF-κB in single and double transgenic mice at the onset stage by Western blot.
As Shc can also be recruited by IGF-1R activatevate downstream SH2-domain containing proteins, we also evaluated the activation of this alternative pathway.
Having established that BV activates the PI3K-Akt signaling axis [1], we next evaluated the activation of pAkt and pS6 (downstream of mTOR) in response to BV in RAW 264.7 macrophages.
We evaluated the activation of the transcription factor, c-jun, by its phosphorylation on an activating residue, Ser63, as well as the electrophoretic shift caused by the phospho-modification of the protein when Western blots were probed with a total anti-c-jun antibody (Fig. 3A and B).
We then evaluated the activation of JNK1/2 (with the use of specific antibodies that recognize the phosphorylated (activated) form of the kinases), and the expression of the anti-apoptotic protein BAK in epithelial tubular cells collected from these patients.
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In the same cell populations, we studied Src activity by the in vitro kinase assay evaluating the activation of Hck, a member of Src kinases.
Next, we performed phospho-kinase arrays, evaluating the activation of key innate immune pathways by HBV antigen and their suppression by HBV infection (Fig. 4a, b).
On the other hand, cyclic voltammetry evaluates the activation of a gold electrode by taking into account the potential difference between the cathodic (E pc) and anodic (E pa) peak currents (ΔE p) (Fig. 1a).
Afterwards, the activated samples with better cleaning performance in terms of resulting NW quality were carefully analyzed by cyclic voltammetry to evaluate the activation of the gold catalyst layer, using it as an indicator of the seed-layer quality for subsequent NW growth.
We used Western blotting analysis to evaluate the activation of ATM in H1299 cells in response to ectopic activation of NPRL2 in the presence or absence of cisplatin (Figure 2A).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com