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ASK1 assays were straightforward and we found ASK1 to be strongly activated by ischaemia in the perfused rat heart, with a gradual increase in activity during ischaemia, reaching maximal activation at 45 min (Fig. 5A).
ERK1 showed a maximal activation at 30 minutes, while ERK2 was barely activated in a time independent way.
Figure 4B demonstrates that BK in A498 cells activates ERK in a concentration-dependent manner, with maximal activation at 1 μM.
Experiments using accelerated argon ions (95 MeV/u, LET ∼230 keV/μm) produced at the French heavy ion accelerator GANIL have shown activation of the NF-κB pathway with doses greater than 1 × 106 particles cm−2 (P cm−2), reaching its maximal activation at 2 × 107 P cm−2.
In initial time course experiments, we found significant phospho-Akt upregulation as early as 2 minutes with maximal activation at 10 15 minutes after exposure of EGNP-1-transduced HUVEC to EGF (Figure 3e).
Over this same range of stretch frequencies, Hosokawa et al. [29] reported a monotonic increase in ERK activation with near maximal activation at 1Hz in vascular smooth muscle cells subjected to 20% cyclic uniaxial stretch.
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4-NP-β-mannoside also showed a higher maximal activation as compared to the corresponding glucoside.
Further addition of the second alpha-helical domain (residues 1-528) did not alter binding to F-actin but resulted in a 2-fold increased activity in promoting adducin binding with half-maximal activation at 50 nM.
Titrating the T3 RNAP template DNA in TX-TL reactions, we found a steep transition from low to high expression with a half-maximal activation at 10 pM (Supplementary Figure 14).
The N-terminal domain (residues 1-313) of beta-spectrin associated with F-actin with a Kd of 26 microM, and promoted adducin binding to F-actin with half-maximal activation at 110 nM.
MVP opens in response to hyperpolarization with half-maximal activation at −175 ± 33 mV and an apparent gating charge of 1.1 e(6).
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