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Cells were exposed to airborne acrolein for 30 min before they were allowed to recover in fresh media, with cell sampling at defined time points to allow evaluation of toxicity and protein damage.
Nevertheless, SPIO labeling suffers from common limitations to other exogenous contrasts, such as dilution of the contrast media with cell division and the possibility that apoptotic stem cells may be phagocytized by macrophages, leading to signal that could be incorrectly associated with the cells.
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High glucose medium contained the same fermentor media (with cells removed), but with glucose added back to a concentration of 1% (w/v).
Cells were washed three times with pre-warmed media, removed with cell dissociation buffer (Gibco) and plate-bound cells were counted by FACS.
After 30 minutes, the media (with ES cells) is removed and cells counted.
Supernatant was aspirated, and cells cultured to confluence in standard media, with nonadherent cells discarded after 5 days.
Co-immunoprecipitation and immunoblotting analysis showed that FGFR4 phosphorylation (tyrosine kinase phosphorylation) was increased in cancer cells maintained in CAF-conditioned media compared with cells incubated in NF- or PF- conditioned media.
The phosphorylation level of Mek/Erk was enhanced in cancer cells maintained in CAF-conditioned media compared with cells incubated in NF- or PF-conditioned media, although no apparent differences were observed in total Mek/Erk expression (Fig. 5f).
Controls consist of cells with media containing DMSO (<0.5%) and blank wells contained media with no cells.
L1 media with no cells was used as a control.
Media with high cell-specific lactate production rates coincided well with high absolute concentrations (Fig. 1c and Table 2).
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