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The cells were immobilized on the window of an observation chamber that was continuously perfused with medium containing exogenous 3OC6HSL autoinducer (AI), so that each cell was subject to a precisely defined local AI concentration.
As expected, for the DAB-treated parasites that were then transferred into medium containing exogenous putrescine, the intermediary TveIF5A isoform was observed with a pI of 5.3.
T. vaginalis was grown in TYM-serum medium (control) or transferred into a medium containing exogenous putrescine after DAB treatment (DAB-treated).
Interestingly, when putrescine biosynthesis was inhibited in T. vaginalis using 1,4-diamino-2-butanone (DAB) and the parasites were then transferred to a medium containing exogenous putrescine, a new TveIF5A spot with a pI of 5.3 was observed [12].
Once a sufficient number of cells had adhered to the window (requiring 15-30 minutes), we supplied autoinducer by connecting the device inputs to syringe pumps that delivered defined medium containing exogenous 3OC6 HSL and/or C8HSL.
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Cells were grown in the estrogen-free medium containing no exogenous compounds for 2 days before transfection.
The correct replacement of the endogenous ERG9 promoter was verified by PCR, and the ERG9-controllable strains exhibited a severe growth defect in medium with DOX; the growth defect caused by DOX was suppressed by the addition of serum containing exogenous cholesterol.
Cells were grown overnight in the same defined medium used for the luminescence experiments and then transferred to fresh medium containing 1000 nM exogenous AI.
We found that esMCs were produced after approximately 18 days of directed differentiation in medium containing just two exogenous growth factors, and esMC continued to be produced for several months when growth factors were replaced.
When grown on 1/2 MS medium containing 10 μM ABA, exogenous ABA inhibited root growth of TaNAC29-overexpression line seedlings more severely than observed in WT and VC, suggesting that TaNAC29 transgenic lines were hypersensitive to ABA (Fig. 4a).
When coronary endothelial cells are incubated in medium containing glucose as the sole exogenous substrate, more than 90% of CO2 production is derived from the PPP, as exogenous glucose is mainly converted into lactate during glycolytic energy production (Krutzfeldt et al, 1990).
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